Team:Heidelberg/Templates/Materials list

From 2013.igem.org

(Difference between revisions)
(Created page with " == Materials used during the project == Partly adopted from iGEM Team Heidelberg 2009 and 2010. Please recheck. '''Table 1: Kits''' {|class="wikitable" |- ! Kit !! Supplier !! ...")
(→Materials used during the project)
Line 168: Line 168:
'''Table 6: Primers and Oligos'''
'''Table 6: Primers and Oligos'''
-
{|class="wikitable"
+
 
 +
==Misc Primers==
 +
 
 +
{| class="wikitable"
|-
|-
-
! Oligo !! Supplier !! Catalog Number
+
! Identifier !! Order date !! Note !! Sequence
|-
|-
-
| BpsA_TE_Stop_8_rbs_6_rv || Sigma-Aldrich Chemie GmbH || VC00021'20637090
+
| VF2 || - || for screening in standard BB backbone- binds on the Backbone before Insert  || tgccacctgacgtctaagaa
|-
|-
-
| pSB1C3_BBa_J04450_rv || Sigma-Aldrich Chemie GmbH || VC00021`20637084
+
| VR || - || for screening in standard BB backbone- binds on Backbone behind Insert || attaccgcctttgagtgagc
|-
|-
-
| pSB1C3_BBa_J04450_fw || Sigma-Aldrich Chemie GmbH || VC00021`20637083
+
|}
 +
 
 +
==Delftibactin==
 +
 
 +
{| class="wikitable"
|-
|-
-
| svp_cds_fw || Sigma-Aldrich Chemie GmbH || VC00021`20637091
+
! Identifier !! Order date !! Note !! Sequence
|-
|-
-
| BpsA_AOxA_rv || Sigma-Aldrich Chemie GmbH || VC00021`20637086
+
| DN01:delH_f1_PacI_fw || - || fw Primer for DelH-fragment1 with RBS and PacI-restriction site || tttt ttaattaa    tcacacaggaaagtactag  ATGGACCGTGGCCGCCTGC    GCCAAATCG
|-
|-
-
| BpsA_TE_fw || Sigma-Aldrich Chemie GmbH || VC00021`20637089
+
| DN02:delH_f1_SalI_rev || - || - || tttt GTCGACCAACACCTGTGCCTGC
|-
|-
-
| BpsA_T_rv || Sigma-Aldrich Chemie GmbH || VC00021`20637088
+
| DN03:delH_f2_SalI_fw || - || - || tttt GTCGACTGGATGGAGCCTGGTGAAAG
|-
|-
-
| BpsA_AOxA_fw || Sigma-Aldrich Chemie GmbH || VC00021`20637085
+
| DN04:delH_f2_KpnI_rev || - || - || tttt ggtacc  TCAGTCCAGCGCGTACTCCAG
|-
|-
-
| BpsA_T_fw || Sigma-Aldrich Chemie GmbH || VC00021`20637087
+
| DN05:AraCbb_KpnI_fw || - || amplifying the Backbone for DelH (pSB6A1-AraC-lacZ) || tttt  ggtacc  aaagaggagaaatactagatgaccatg
|-
|-
-
| svp_cds_rv || Sigma-Aldrich Chemie GmbH || VC00021`20637092
+
| DN06:AraCbb_PacI_rev2 || 15-05-2013  || amplifying the Backbone for DelH (pSB6A1-AraC-lacZ) || tttt  ttaattaa  gctagcccaaaaaaacggtatg
|-
|-
-
| IK36:pLF03_seq_rev || Sigma-Aldrich Chemie GmbH || VC00021`21393236
+
| DN07:Screen_delH_rev|| 15-05-2013 || for screening if DelH is present - binds on the very beginning of DelH || CTTTCCTCGAACACCGTGCGCAG
|-
|-
-
| IK37:pLF03-catR_seq_rev || Sigma-Aldrich Chemie GmbH || VC00021`21393237
+
| DN08:DelH_EcoRI_rev || - || rev_Primer for DelH Fragment f1a || CTCGTCGCCATGGACCAGGCAG
|-
|-
-
| SR_01_FS34_fwtoFS26 || Sigma-Aldrich Chemie GmbH || VC00021`21457452
+
| DN09:DelH_f1_fw_long || 2013-06-11 || for amplifying DelH-1a from the genome: doesn't work|| ATGGACCGTGGCCGCCTGCGCCAAATCG
|-
|-
-
| FS_27_OP_rv || Sigma-Aldrich Chemie GmbH || VC00021`21457453
+
| DN10:DelH_f1_fw_short || 2013-06-11 || for amplifying DelH-1a from the genome: doesn't work || ATGGACCGTGGCCGCCTGC
|-
|-
-
| FS_28_OP_rv || Sigma-Aldrich Chemie GmbH || VC00021`21457454
+
| DN11:DelH_f1_fw_short2|| 2013-06-11 || for amplifying DelH-1a from the genome: works!!!! || GCCGCCTGCGCCAAATCG
|-
|-
-
| FS_29_OP_rv || Sigma-Aldrich Chemie GmbH || VC00021`21457455
+
| DN12:DelH_f1_PacI_fw_short|| 2013-06-11 || for amplifying DelH-1a from the genome: doesn't work || ttttttaattaatcacacaggaaagtactagATGGACCGTGGCCGCCTGC
|-
|-
-
| FS_30_OP_rv || Sigma-Aldrich Chemie GmbH || VC00021`21457456
+
| DN13:Screen_DelH_fw|| 15-05-2013 || PCR Screening for presence of DelH insert || gtaaacccactggtgataccattc
|-
|-
-
| FS_31_OP_fw || Sigma-Aldrich Chemie GmbH || VC00021`21457457
+
| FS_01: pSB4K5_DelA_rv||20-13-06-28||Amplification of pSB4K5 from the iGEM Distribution Gibson Primer with overhang to DelA introducing the RBS BBa_B0035|| TCGCGGCGATCCGGTACTGCGCCTCTGTTGAACATCTGATATTCTCCTCTTTAATCGACAGATTGTGTGAAATTGTTATCCGCTCAC
|-
|-
-
| FS_32_OPgood_fw || Sigma-Aldrich Chemie GmbH || VC00021`21457458
+
|FS_02: DelAG_1_fw||2013-06-28 || Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| TTCAACAGAGGCGCAGTACCGGATC
|-
|-
-
| FS_33_OP_fw || Sigma-Aldrich Chemie GmbH || VC00021`21457459
+
|FS_03: DelAG_1_rv||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| GTCGGAGACGATGTGGTGCATCAC
|-
|-
-
| RB46:indC-woPPT_rv || Sigma-Aldrich Chemie GmbH || VC00021`21487465
+
|FS_04: DelAG_2_fw||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| CTGCAGGCCAATGAGCACATCCTG
|-
|-
-
| RB47:indC-SpeI_fw || Sigma-Aldrich Chemie GmbH || VC00021`21487466
+
|FS_05: DelAG_2_rv||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| CACAGGTGGTAGATGGCGTC
|-
|-
-
| RB48:indC-EcoRI_rv || Sigma-Aldrich Chemie GmbH || VC00021`21487467
+
|FS_06: DelAG_3_fwG||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| ATTGCGAGGACTTGCTCGATG
|-
|-
-
| RB49:indC-EcoRI_fw || Sigma-Aldrich Chemie GmbH || VC00021`21487468
+
|FS_07: DelAG_3_rv||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| TTTGCTGCAGCGCCAGCACATCGAG
|-
|-
-
| RB50:indC-SpeI_rv || Sigma-Aldrich Chemie GmbH || VC00021`21487469
+
|FS_08: DelAG_4_fw||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| GTACGGCCTATCACATCAGCG
|-
|-
-
| RB51:pSB(2/3)K3_fw || Sigma-Aldrich Chemie GmbH || VC00021`21487470
+
|FS_09: DelAG_4_rv||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| GAAGCTCAGCAGGTTGGGCGAGACG
|-
|-
-
| RB52:pSB(2/3)K3_rv || Sigma-Aldrich Chemie GmbH || VC00021`21487471
+
|FS_10: DelAG_5_fw||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| GAATTTTGTTCCACCACCTGCTG
|-
|-
-
| RB57:tycC6-T_fw || Sigma-Aldrich Chemie GmbH || VC00021`21522074
+
|FS_11: DelAG_5_rv||2013-06-28 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer with overhang to DelOP|| CTTGAGCAGGCGCAGTACCTCGGAGGGCGGTCGGCTGGCGTTTTCCATGATTCAGGTTTCCTGTGTGAAGCTCATCTCAGATATCTCCCAGAGTTTCGAGAAAG
|-
|-
-
| RB58:tycC6-T_rv || Sigma-Aldrich Chemie GmbH || VC00021`21522075
+
|FS_11: DelAG_5_short_rv||2013-05-07 ||Amplification of DelAG from ''Delftia acidovorans'' genome Gibson Primer|| TCAGATATCTCCCAGAGTTTCGAGAAAG
|-
|-
-
| RB56:tycA1-T_rv || Sigma-Aldrich Chemie GmbH || VC00021`21522073
+
|FS_12: DelOP_fw||2013-06-28 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer|| GAATCATGGAAAACGCCAGCCGAC
|-
|-
-
| RB55:tycA1-T_fw || Sigma-Aldrich Chemie GmbH || VC00021`21522072
+
|FS_13: DelOP_rv||2013-06-28 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer with overhang to DelL|| CAATGTTGGAGGGGCCGAAGCCGATGCCGATCAGCGGGTGGGTTTGCATGGAAGGTCCTTTCATTGGGTCGATGCGTCCAGTGTCACACCGTGGTGTCTGCAGGCG
|-
|-
-
| RB53:entF-T_fw || Sigma-Aldrich Chemie GmbH || VC00021`21522070
+
|FS_13: DelOP_short_rv||2013-05-07 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer with overhang to DelL||TCACACCGTGGTGTCTGCAGGCG
|-
|-
-
| RB54:entF-T_rv || Sigma-Aldrich Chemie GmbH || VC00021`21522071
+
|FS_14: DelL_fw||2013-06-28 ||Amplification of DelL from ''Delftia acidovorans'' genome Gibson Primer|| CAAACCCACCCGCTGATCGGCATC
|-
|-
-
| RB67:DelC-pSB2K3_rv || Sigma-Aldrich Chemie GmbH || VC00021`21522084
+
|FS_15: DelL_mRFP_pSB4K5_rv||2013-06-28 ||Amplification of DelL Gibson Primer with overhang to BBa_J04450|| GAAACGCATGAACTCTTTGATAACGTCTTCGGAGGAAGCCATCTAGTATTTCTCCTCTTTCTCTAGTATCAGTCCTGCAGCGCCAGCTGTTCTGTG
|-
|-
-
| RB66:DelC-pSB2K3_fw || Sigma-Aldrich Chemie GmbH || VC00021`21522083
+
|FS_15: DelL_mRFP_pSB4K5__short_rv||2013-05-07 ||Amplification of DelL Gibson Primer with overhang to BBa_J04450||TCAGTCCTGCAGCGCCAGCTGTTCTGTG
|-
|-
-
| RB63:pSB(2/3)K3_rv_korr || Sigma-Aldrich Chemie GmbH || VC00021`21522080
+
|FS_16: mRFP_pSB4K5_fw (Del)||2013-06-28 ||Amplification of pSB4K5 from iGEM Distribution Gibson Primer|| GCTTCCTCCGAAGACGTTATC
|-
|-
-
| RB65:DelC-extr_rv || Sigma-Aldrich Chemie GmbH || VC00021`21522082
+
|FS_20: DelF_fw||2013-07-13 ||Amplification of DelF from ''Delftia acidovorans'' genome Gibson Primer|| GACTTGCTCGATGCGGTGCAG
|-
|-
-
| RB59:delH4-T_fw || Sigma-Aldrich Chemie GmbH || VC00021`21522076
+
|FS_21: DelF_fw||2013-07-13 ||Amplification of DelF from ''Delftia acidovorans'' genome Gibson Primer|| GACGCCATCTACCACCTGTG
|-
|-
-
| RB60:delH4-T_rv || Sigma-Aldrich Chemie GmbH || VC00021`21522077
+
|FS_22: DelOP_short_fw||2013-08-02 ||Amplification of DelOP from ''Delftia acidovorans'' genome inlcuding the recently predicted endogenous Promotor for DelOP Gibson Primer|| GATGACGCAGGGCGGCGGAATTTGTTCATC
|-
|-
-
| RB61:delH5-T_fw || Sigma-Aldrich Chemie GmbH || VC00021`21522078
+
|FS_23: DelG_long_rv||2013-07-13 ||Amplification of DelG from ''Delftia acidovorans'' genome Gibson primer with overhang to DelOP element including the recently predicted endogenous Promotor|| GATGAACAAATTCCGCCGCCCTGCGTCATCTCAGATATCTCCCAGAGTTTCGAGAAAG
|-
|-
-
| RB62:delH5-T_rv || Sigma-Aldrich Chemie GmbH || VC00021`21522079
+
|FS_24: DelAE_rv||2013-07-13 ||Amplification of DelAE from ''Delftia acidovorans'' genome Gibson Primer|| CAGAAGAATTCCCAGAAGGAGATGTCGAAG
-
|-  
+
-
| RB64:DelC-extr_fw || Sigma-Aldrich Chemie GmbH || VC00021`21522081
+
|-
|-
-
| KH9:ccdB-Big_fw || Sigma-Aldrich Chemie GmbH || VC00021`21522085
+
|FS_25: DelEF_fw||2013-07-13 ||Amplification of DelEF from ''Delftia acidovorans'' genome Gibson Primer|| ACACGGTGCTGCAGAAAACGCCCTTC
|-
|-
-
| KH10:ccdB-Big_rv || Sigma-Aldrich Chemie GmbH || VC00021`21522086
+
|FS_26: DelFG_rv||2013-07-13 ||Amplification of DelFG from ''Delftia acidovorans'' genome Gibson Primer|| GAATTCATCCACGATGATCTGCATG
|-
|-
-
| AT08:RFC10suffix_TycC6_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564137
+
|FS_27: DelOP_rv||2013-08-02 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer|| CTTTGGGCCGTGCCGGTTTTTGAGATAC
|-
|-
-
| KH13_BBa-entD_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564138
+
|FS_28: DelOP_rv||2013-08-02 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer|| GTTTTTGAGATACGCGCGTTGTCAC
|-
|-
-
| AT03:RFC10prefix-TycB1_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564132
+
|FS_29: DelOP_rv||2013-08-02 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer|| TTCCCCCTCTCTTTCTCGCTTC
|-
|-
-
| AT06:RFC10suffix_TycC5_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564135
+
|FS_30: DelOP_rv||2013-08-02 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer|| CCGCTTCCCCCTCTCTTTCTCGCTTC
|-
|-
-
| AT02:RFC10suffix_TycA_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564131
+
|FS_31: DelOP_fw||2013-08-02 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer|| GTTGGCGAGTTCAAGAAATG
|-
|-
-
| AT01:RFC10prefix_TycA_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564130
+
|FS_32: DelOP_fw||2013-08-02 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer|| TCCTTCAGGTGTGCGGCAGACAAG
|-
|-
-
| AT05:RFC10prefix-TycC5_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564134
+
|FS_33: DelOP_fw||2013-08-02 ||Amplification of DelOP from ''Delftia acidovorans'' genome Gibson Primer|| TTCTTCGTGATGACGCAGGGCGGCGGAATTTGTTC
|-
|-
-
| AT07:RFC10prefix-TycC6_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564136
+
|FS_35: DelG_fw||2013-08-05 ||Amplification of DelG from ''Delftia acidovorans'' genome Gibson Primer|| CATGCAGATCATCGTGGATGAATTC
|-
|-
-
| AT04:RFC10suffix_TycB1_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564133
+
|FS_45: pSB4K5_fw||2013-08-02 ||Amplification of pSB4K5 from iGEM Distribution Gibson Primer without mRFP|| CCAGGCATCAAATAAAACGAAAG
|-
|-
-
| KH21_BBa-indC_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564146
+
|FS_46: DelL_rv ||2013-08-02 ||Amplification of DelL from ''Delftia acidovorans'' genome Gibson Primer creating overlap to pSB4K5 without mRFP|| TCAGTCCTGCAGCGCCAGCTGTTCTGTGCTTTCGTTTTATTTGATGCCT
|-
|-
-
| KH20_BBa-delC_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564145
+
|FS_47_screening_BB_AF_fw||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| GTTGGCCGATTCATTAATGC
|-
|-
-
| KH22_BBa-indC_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564147
+
|FS_48_screening_BB_AF_rv||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| TAACGGTATCGGTATCGCTTTG
|-
|-
-
| KH15_BBa-sfp_Bsub_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564140
+
|FS_49_screening_AFI_AFII_fw||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| GTTTCTCTGGAAGATGGATAC
|-
|-
-
| KH16_BBa-sfp_Bsub_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564141
+
|FS_50_screening_AFI_AFII_rv||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| GTTGACGAAAAAGCCGACCAC
|-
|-
-
| KH17_BBa-svp_Svert_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564142
+
|FS_51_screening_AF_FG(21-26)_fw||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| TGGATATCGACTGGACTGCCTG
|-
|-
-
| KH14_BBa-entD_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564139
+
|FS_52_screening_AF_FG(21-26)_rv||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| TGCACCACATCGACGAAACGG
|-
|-
-
| KH18_BBa-svp_Svert_rv || Sigma-Aldrich Chemie GmbH || VC00021`21564143
+
|FS_53_screening_FG(21-26)_G_fw||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| GTACGGCCTATCACATCAGCG
|-
|-
-
| KH19_BBa-delC_fw || Sigma-Aldrich Chemie GmbH || VC00021`21564144
+
|FS_54_screening_FG(21-26)_G_rv||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| GAACCTGGGTGTTCACGAAAAAGCC
|-
|-
-
| KH13_BBa-entD_fw || Sigma-Aldrich Chemie GmbH || VC00021`21566979
+
|FS_55_screening_G_OP_fw||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| GTATCTCTACATGCATCGCTAC
|-
|-
-
| KH14_BBa-entD_rv || Sigma-Aldrich Chemie GmbH || VC00021`21566980
+
|FS_56_screening_G_OP_fw||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| AGGACATTTTCCGCACCCCG
|-
|-
-
| NK_01_FS_62_screen_nomRFP_rv || Sigma-Aldrich Chemie GmbH || VC00021`21599362
+
|FS_57_screening_G_OP_rv||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| GCTGGCGTTTTCCATAAG
|-
|-
-
| FS_46_L_rv_gibsontoBBpSB4K5 || Sigma-Aldrich Chemie GmbH || VC00021`21599355
+
|FS_58_screening_OP_L_fw||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| GAACAACTTCCAGCACAGCCTGTTC
|-
|-
-
| FS_61_screening_with_mRFP || Sigma-Aldrich Chemie GmbH || VC00021`21599361
+
|FS_59_screening_OP_L_rv||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| CGTTGAAGATTTCGTTGACG
|-
|-
-
| FS_52_screening_AF_FG21_26_rv || Sigma-Aldrich Chemie GmbH || VC00021`21599359
+
|FS_60_screening_L_BB_fw||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| CATCTTCAAGGTGTTCTATGAAC
|-
|-
-
| FS_51_screening_AF_FG21_26_fw || Sigma-Aldrich Chemie GmbH || VC00021`21599358
+
|FS_61_screening_L_BB(with_mRFP)_rv||2013-08-02 ||Primer for screening/sequencing of pFSN construct|| CAGTTTAACTTTGTAGATGAAC
|-
|-
-
| FS_45_pSB4K5_fw_wo_mRFP || Sigma-Aldrich Chemie GmbH || VC00021`21599354
+
|FS_66: DelH_rv||2013-08-26 || Amplification of DelH from ''Delftia acidovorans'' genome Gibson Primer|| TGGGCATTCACCGCATCGATC
|-
|-
-
| FS_60_screening_L_BB_fw || Sigma-Aldrich Chemie GmbH || VC00021`21599360
+
|FS_67: DelH_fw||2013-08-26 || Amplification of DelH from ''Delftia acidovorans'' genome Gibson Primer|| CTTCACGTTGATTGCGCATG
|-
|-
-
| FS_48_screening_BB_AF_rv || Sigma-Aldrich Chemie GmbH || VC00021`21599357
+
|FS_68: DelH_rv||2013-08-26 || Amplification of DelH from ''Delftia acidovorans'' genome Gibson Primer|| CAGAAGAACTCCCAGACCGAC
|-
|-
-
| FS_47_screening_BB_AF_fw || Sigma-Aldrich Chemie GmbH || VC00021`21599356
+
|FS_69: DelH_fw||2013-08-26 || Amplification of DelH from ''Delftia acidovorans'' genome Gibson Primer|| GACACCGTTCAGCTTCGATG
|-
|-
-
| AT05_RFC10prefix_TycC5_fw || Sigma-Aldrich Chemie GmbH || VC00021`21599363
+
|FS_70: DelH_rv||2013-08-26 || Amplification of DelH from ''Delftia acidovorans'' genome Gibson Primer|| GAAGCTGCTCCGCTGATAGAT
|-
|-
-
| AT07_RFC10prefix_TycC6_fw || Sigma-Aldrich Chemie GmbH || VC00021`21599364
+
|FS_71: DelH_fw||2013-08-26 || Amplification of DelH from ''Delftia acidovorans'' genome Gibson Primer|| ATGTGCTGTCGCTCAAGATG
|-
|-
-
| PW17:TycAdE-C(TycC2)_rev || Sigma-Aldrich Chemie GmbH || VC00021`21632288
+
|FS_72_SR_02_fw||2013-08-30 || Screening of pFHFSN|| ATGTGCTGTCGCTCAAGATG
|-
|-
-
| PW15:C(TycC2)-indC_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21632286
+
|FS_73_SR_03_fw||2013-08-30 || Screening of pFHFSN|| GTGCTGTTTGGCCGTATG
|-
|-
-
| PW14:C(TycC2)-indC_rev || Sigma-Aldrich Chemie GmbH || VC00021`21632285
+
|FS_74_SR_04_fw||2013-08-30 || Screening of pFHFSN|| ATCAGGTGCTGAGCTACGAC
|-
|-
-
| PW16:indC_rev || Sigma-Aldrich Chemie GmbH || VC00021`21632287
+
|FS_75_SR_05_fw||2013-08-30 || Screening of pFHFSN|| CTGTTCATCAACACCTTGCC
|-
|-
-
| KH11_plu2642-T_fw || Sigma-Aldrich Chemie GmbH || VC00021`21632281
+
|FS_76_SR_06_rv||2013-08-30 || Screening of pFHFSN|| GAAGACAGTCATAAGTGCGGC
|-
|-
-
| RB77:plu2670-T_rv || Sigma-Aldrich Chemie GmbH || VC00021`21632284
+
|FS_77_rv|| 2013-09-11 ||Gibson-Primer rev, Amplficiation of the Backbone pSB6A1 with overlap to the RBS BBa_B0034 and the lacI-promotor, it creates an overlap to the beginning of DelH|| GCGATTTGGCGCAGGCGGCCACGGTCCATCTAGTATTTCTCCTCTTTC
|-
|-
-
| KH12_plu2642-T_rv || Sigma-Aldrich Chemie GmbH || VC00021`21632282
+
|FS_78_rv|| 2013-09-26 ||Gibson-Primer rev, Amplficiation of the Backbone pSB6A1 introducing the RBS BBa_B0032 and the promotor BBa_J23114  and creating an overlap to the first fragment of DelH amplified with primer DN_11|| GATTTGGCGCAGGCGGCCACGGTCCATCTAGTACTTTCCTGTGTGACTCTAGAGCTAGCATTGTACCTAGGACTGAGCTAGCCATAAACTCTAGAAGCGGCCGCGAATTC
|-
|-
-
| RB76:plu2670-T_fw || Sigma-Aldrich Chemie GmbH || VC00021`21632283
+
|FS_84_fw|| 2013-09-26 ||Gibson-Primer fw, Amplficiation of the first fragment of DelH introducing the RBS BBa_B0032 and creating an overlap to primer FS_85 thereby partially introducing the promotor BBa_J23114|| GCTCAGTCCTAGGTACAATGCTAGCTCTAGAGTCACACAGGAAAGTACTAGATGGACCGTGGCCGCCTGCG
|-
|-
-
| PW18:TycAdE-C(TycC2)_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21632289
+
|FS_85_rv|| 2013-09-26 ||Gibson-Primer rev, Amplficiation of the Backbone pSB6A1, partially introducing the promotor BBa_B0032 with overlap to primer FS_84 and therefore the promotor BBa_J23114, it creates an overlap to the beginning of DelH|| GCGATTTGGCGCAGGCGGCCACGGTCCATCTAGTATTTCTCCTCTTTC
|-
|-
-
| PW19:TycC4dC_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21632290
+
|FS_86_rv|| 2013-09-27 ||Gibson-Primer rev, Amplficiation of the Backbone pSB4K5 without any promotor introducing a KpnI cutting site for restriction cloning, creates an overlap to DelH and will be used for the ccdB strategy|| GGCGATTTGGCGCAGGCGGCCACGGTCCATGTACTTCGAGTCACTAAGGGCTAAC
|-
|-
-
| PW20:TycC4dC-C(TycC2)_rev || Sigma-Aldrich Chemie GmbH || VC00021`21632291
+
|FS_87_fw|| 2013-09-27 ||Gibson-Primer fw, Amplficiation of the Backbone pSB6A1 introducing a BamHI cutting site for restriction cloning and creating an overlap to the last fragment of DelH|| CGCTGGAGTACGCGCTGGACTGAGATCCCAGGCATCAAATAAAACG
|-
|-
-
| PW21:TycC4dC-C(TycC2)_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21632292
+
|FS_90_fw|| 2013-09-27 ||Gibson-Primer fw, Amplficiation of the ccdB cassette from the template pDonorPlasmid introducing a KpnI cutting site for restriction cloning, creates an overlap to the promotor BBa_J23114 and will be used for the ccdB strategy|| CTCAGTCCTAGGTACAATGCTAGCTCTAGAGTCACACAGGAAAGCAGTACACTGGCTGTGTATAAGGGAG
|-
|-
-
| PW22:pSB1C3-TycC4dC_rev || Sigma-Aldrich Chemie GmbH || VC00021`21632293
+
|FS_93_rv|| 2013-09-27 ||Gibson-Primer rev, Amplficiation of the ccdB cassette from the template pDonorPlasmid introducing a BamHI cutting site for restriction cloning, creates an overlap to the backbone pSB6A1 and will be used for the ccdB strategy|| GTTCACCGACAAACAACAGATGATCCGCGTGGATCCGGCTTAC
|-
|-
-
| PW23:indC-pSB1C3_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21632294
+
|FS_94_fw|| 2013-09-27 ||Primer fw, Amplficiation of the backbone pSB6A1, will be used for the ccdB strategy|| ATCTGTTGTTTGTCGGTGAACGC
|-
|-
-
| PW24:TycC1dC_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21632295
+
| HM01:DelH_EcoRI_fw || - || fw_Primer for DelH Fragment f1b|| GCATTGGAGCCTCAATGGCAAGTC
|-
|-
-
| PW25:pSB1C3-TycC1dC || Sigma-Aldrich Chemie GmbH || VC00021`21632296
+
|HM02:DelH_Gib1.1_rev||2013-07-09 ||Gibson-Primer DelH|| TGCTGCGCCTGCATACGGCCAAACA
|-
|-
-
| HM13:Screen_DelH_end_fw || Sigma-Aldrich Chemie GmbH || VC00021`21635842
+
|HM03:DelH_Gib1.2_fw||2013-07-09 ||Gibson-Primer DelH|| AGCGGCAGGGACGACGTGGT
|-
|-
-
| HM14:DelH_tetR_fw || Sigma-Aldrich Chemie GmbH || VC00021`21635843
+
|HM04:DelH_Gib1.2_rev||2013-07-09 ||Gibson-Primer DelH|| CATAGAGGTTGTAGAGA
|-
|-
-
| HM15:tetR_stop_BB_rev || Sigma-Aldrich Chemie GmbH || VC00021`21635844
+
|HM05:DelH_Gib2.1_fw||2013-07-09 || Gibson-Primer DelH || AGAACGCCGTCTTCAGGCTCCTG
|-
|-
-
| HM16:tetR_pSB6A1_fw || Sigma-Aldrich Chemie GmbH || VC00021`21635845
+
|HM06:DelH_Gib2.1_rev||2013-07-09 ||Gibson-Primer DelH|| CAATGCTTTG CCGCTCGAA
|-
|-
-
| HM17:DelH_Terminator_BB_fw || Sigma-Aldrich Chemie GmbH || VC00021`21635846
+
|HM07:DelH_Gib2.2_fw||2013-07-09 ||Gibson-Primer DelH|| TCGCCACGGCAGCTGTTCGA
|-
|-
-
| FS_63_rv_scre_mRFP_to_DelL || Sigma-Aldrich Chemie GmbH || VC00021'21714285
+
|HM08:DelH_Gib2_end_rev||2013-07-09 ||Gibson-Primer DelH|| TCAGTCCAGCGCGTACTCCAG
|-
|-
-
| FS_57_rv_scre_DelOP_to_DelG || Sigma-Aldrich Chemie GmbH || VC00021'21714286
+
|HM09:AraC_RBS_Delh_rev||2013-07-09||Gibson-Primer rev, introduces a new RBS and has the AraC-promotor and the beginning of DelH|| TTGCAAAGCGCTCGGCGATTTGGCGCAGGCGGCCACGGTCCATttaactTTCTCCTCTTTAATactttgagctagcccaa
 +
Aaaaacggtatggagaaacagtagagagtt
|-
|-
-
| FS_58_fw_scre_DelOP_to_DelL || Sigma-Aldrich Chemie GmbH || VC00021'21714287
+
|HM10:RBS_lacZ||2013-07-09 ||Gibson-Primer fw for the pSB6A1 Backbone with the end of DelH, RBS(1) and the beginning of lacZ || TGGAGTACGCGCTGGACTGA  TCTAGAG  AAAGAGGAGAAA TACTAG ATGACCATGATTA
|-
|-
-
| AT09_R10_B1_fw_longer || Sigma-Aldrich Chemie GmbH || VC00021`21736751
+
|HM11:lacI_RBS(1)_DelH_rev|| 2013-07-24 ||Gibson-Primer rev, amplify the Backbone with overlap with the RBS and the lacI-promotor and it creates and overlap to the start of DelH|| TCGGCGATTTGGCGCAGGCGGCCACGGTCCATctagtatttctcctctttctctagtatgtgtg
|-
|-
-
| AT10_R10_B1_rv_longer || Sigma-Aldrich Chemie GmbH || VC00021`21736752
+
|HM12:DelH_RBS(1.2)_mRFP_fw|| 2013-07-24 || Gibson-Primer fw for the pSB6A1 Backbone with the end of DelH, introducing a new RBS(new) and the beginning of mRFP|| ATTGGCGCTGGAGTACGCGCTGGACTGAtcaaagtATTAAAGAGGAGAAagttaaatggcttcctccgaagacgttatcaAagag
|-
|-
-
| AT11_R10_C5_fw_rpos || Sigma-Aldrich Chemie GmbH || VC00021`21736753
+
|HM13:Screen_DelH_end_fw||2013-08-16 ||New screening primer for the end of DelH together with the VR2 primer from the registry|| TTTCTGACGACCCTGCACCTGAAG
|-
|-
-
| RB78_indC_SpeI_rv_end || Sigma-Aldrich Chemie GmbH || VC00021`21736755
+
|HM14:DelH_tetR_fw||2013-08-16 ||Gibson-Primer DelH-tetR: amplifies the tetracycline resistance from the pSB1T3 Backbone and creates an overlap to the end of DelH|| ATTGGCGCTGGAGTACGCGCTGGACTGA    atgaagttttaaatcaatctaaag
|-
|-
-
| FS_66_DelH4_rv || Sigma-Aldrich Chemie GmbH || VC00021`21736757
+
|HM15:tetR_stop_BB_rev||2013-08-16 ||Gibson-Primer tetR-pSB6A1: amplifies the tetracycline resistance and creates an overlap with the Terminator of the Backbone pSB6A1|| Cgactgagcctttcgttttatttgatgcctggc  ctcgtgatacgcctatttttatagg
|-
|-
-
| FS_68_DelH6_rv || Sigma-Aldrich Chemie GmbH || VC00021`21736759
+
|HM16:tetR_pSB6A1_fw||2013-08-16||Gibson-Primer DelH, amplifies the Backbone pSB6A1 creating an overlap with the tetracycline resistance || Aaaaataggcgtatcacgag  gccaggcatcaaataaaacgaaaggctcag
|-
|-
-
| FS_67_DelH5_fw || Sigma-Aldrich Chemie GmbH || VC00021`21736758
+
|HM17:DelH_Terminator_BB_fw|| 2013-08-16 || Gibson-Primer fw for the pSB6A1 Backbone (binding the terminator) and creating an overlap with the end of DelH|| ATTGGCGCTGGAGTACGCGCTGGACTGA  aggcatcaaataaaacgaaaggctcag
|-
|-
-
| FS_65_DelH3_fw || Sigma-Aldrich Chemie GmbH || VC00021`21736756
+
| HM20:BB_HPLC_rev || 11-09-2013 || HPLC version of HM11 <br/> Gibson-Primer rev, amplify the Backbone with overlap with the <br/>RBS and the lacI-promotor and it creates and overlap to <br/>the start of DelH  || GATTTGGCGCAGGCGGCCACGGTCCATctagtatttctcctctttc
|-
|-
-
| FS_64_DelH2_rv || Sigma-Aldrich Chemie GmbH || VC00021`21736754
+
| HM21:fw_lacI_BbsI_Xba || 2013-09-15 || Forward primer for cutting out mutated fragment for mutagenesis || TTTTGAAGACAA CTAGGCAATACGCAA
|-
|-
-
| FS_69_DelH7_fw || Sigma-Aldrich Chemie GmbH || VC00021`21736760
+
| HM22:rev_RBS || 2013-09-15 || Reverse Primer in RBS for mutagenesis || TTTTGAAGACAA CTCTTTCTCTAGTATGTGTGAAATTG
|-
|-
-
| FS_70_DelH8_rv || Sigma-Aldrich Chemie GmbH || VC00021`21736761
+
| HM23:fw_RBS || 2013-09-15 || Forward Primer in RBS for mutagenesis || TTTTGAAGACAA AGAGGAGAAATACTAGATGGACCGTGGC
|-
|-
-
| FS_71_NK_02_DelH9_fw || Sigma-Aldrich Chemie GmbH || VC00021`21736762
+
| HM24:rev_BbsI_MfeI || 2013-09-15 || Reverse primer for cutting out mutated fragment for mutagenesis || TTTTGAAGACAA AATTGGACAGCGCGGCATGCCGGTTG
|-
|-
-
| RB79_indC_woPPTase_rv2 || Sigma-Aldrich Chemie GmbH || VC00021`21784357
+
| IK01:pLF03_integr_argK_fw || 2013-06-12 || For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome. || GCTGATGGAAGTGGCTGATCTGATC
|-
|-
-
| IK38:pIK1_fw || Sigma-Aldrich Chemie GmbH || VC00021`21792233
+
| IK02:pLF03_integr_pET21c_rev || 2013-06-12 || For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against pLF03 backbone (pET-21c). || TCCGCTCACAATTCCCCTATAGTG
|-
|-
-
| IK39:pIK1_rev || Sigma-Aldrich Chemie GmbH || VC00021`21792234
+
| IK03:pLF03_integr_argK_rev || 2013-06-18 || For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome for positive control (to be used with IK01 or IK04). || GATAAATTCACTGAGCTGCCGCAG
|-
|-
-
| FS_76_SR_06_rv || Sigma-Aldrich Chemie GmbH || VC00021`21806264
+
| IK04:pLF03_integr_argK_fw || 2013-06-18 || For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome. Alternative forward primer in case IK01 does not work. || GCGGGAATTAATGCTGTTATGCGAAG
|-
|-
-
| FS_72_SR_02_fw || Sigma-Aldrich Chemie GmbH || VC00021`21806265
+
| IK05:pLF03_integr_ygfG_fw || 2013-06-18 || For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome, namely ygfG, which should be replaced by the construct. || GCGGTCATTGAGTTTAACTATGGCC
|-
|-
-
| FS_73_SR_03_fw || Sigma-Aldrich Chemie GmbH || VC00021`21806266
+
| IK06:pLF03_integr_ygfG_rev || 2013-06-18 || For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome, namely ygfG, which should be replaced by the construct. || AACATGGTTGAGGATGCCGACAGC
|-
|-
-
| FS_74_SR_04_fw || Sigma-Aldrich Chemie GmbH || VC00021`21806267
+
| IK07:ygfG21C1 || 2013-06-25 || For insertion of methylmalonyl-CoA synthesis pathway into ''E. coli''. Extended ygfG21C1 with higher melting temperature. || TCACCGCGCACCGGCCTGCGGCAGCTCAGTGAATTTATCC AGATCTCGATCCCGCGAAATTAATAC
|-
|-
-
| FS_75_SR_05_fw || Sigma-Aldrich Chemie GmbH || VC00021`21806268
+
| IK08:ygfG21C2 || 2013-06-25 || For insertion of methylmalonyl-CoA synthesis pathway into ''E. coli''. Extended ygfG21C2 with higher melting temperature. || GTTATGCTGGATAATTTCTGCCGCTTCATTGGCGGTCATC CAAAAAACCCCTCAAGACCCGTTTAG
|-
|-
-
| PW33:TycC6dTE-C(TycC2)_rev || Sigma-Aldrich Chemie GmbH || VC00021`21841357
+
| IK24:pccB_fw || 2013-07-16 || Colony-PCR of methylmalonyl-CoA synthesis pathway || CGTATCGAGGAAGCGACGCAC
|-
|-
-
| PW34:TycC6dTE-C(TycC2)_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21841358
+
| IK25:pccB_rev || 2013-07-16 || Colony-PCR of methylmalonyl-CoA synthesis pathway || GGTGATGAACATGTGGCTGGTCTG
|-
|-
-
| PW27:TycC5dC-C(TycC4)_rev || Sigma-Aldrich Chemie GmbH || VC00021`21841351
+
| IK26:BBa_I746200_fw || 2013-07-19 || Forward primer for permeability device || ATGAACAAGAAGATTCATTCCCTGGCCTTG
|-
|-
-
| PW28:TycC5dC-C(TycC4)_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21841352
+
| IK27:BBa_I746200-BBa_B0029_rev || 2013-07-19 || Reverse primer for permeability device with partial Gibson overhang for BBa_B0029 RBS || GAATACCAGT TCAGAAGTGGGTGTTTACGCTCATATAC
|-
|-
-
| PW29:C(TycC4)-TycC4dC_rev || Sigma-Aldrich Chemie GmbH || VC00021`21841353
+
| IK28:pccB-BBa_B0029-BBa_I746200_fw || 2013-07-19 || Forward primer for pccB with BBa_B0029 RBS and Gibson overhang for permeability device || ATATGAGCGTAAACACCCACTTCTGAACTGGTATTCACACAGGAAACCTACTAG ATGTCCGAGCCGGAAGAGC
|-
|-
-
| PW30:C(TycC4)-TycC4dC_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21841354
+
| IK29:accA2-BBa_B0030_rev || 2013-07-19 || Reverse primer for accA2 with partial Gibson overhang for BBa_B0030 RBS || TAATGAAGTTG CTAGTGATTCTCGCAGATGGC
|-
|-
-
| PW31:TycC4-C(TycC4)_rev || Sigma-Aldrich Chemie GmbH || VC00021`21841355
+
| IK30:sfp-BBa_B0030-accA2_fw || 2013-07-19 || Forward primer for sfp with BBa_B0030 RBS and Gibson overhang for accA2 || TCCGGCGCCGCCATCTGCGAGAATCACTAGCAACTTCATTAAAGAGGAGAAATACTAG ATGAAGATTTACGGAATTTATATGGAC
|-
|-
-
| PW32:TycC4-C(TycC4)_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21841356
+
| IK31:sfp_rev || 2013-07-19 || Reverse primer for sfp
|-
|-
-
| PW26:TycC5dC_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21841350
+
| IK32:BBa_J04450-sfp_fw || 2013-07-19 || Forward primer for mRFP-containing backbones with Gibson overhang for sfp || ATCACAATGGTCTCGTACGAAGAGCTTTTATAA TACTAGAGCCAGGCATCAAATAAAACG
|-
|-
-
| PW35:TycC6dTE-C(TycC4)_rev || Sigma-Aldrich Chemie GmbH || VC00021`21841359
+
| IK33:BBa_J04450-BBa_I746200_rev || 2013-07-19 || Reverse primer for mRFP-containing backbones with Gibson overhang for permeability device || ACAAGGCCAGGGAATGAATCTTCTTGTTCAT CTAGTATTTCTCCTCTTTCTCTAGTATG
|-
|-
-
| PW36:TycC6dTE-C(TycC4)_fwd || Sigma-Aldrich Chemie GmbH || VC00021`21841360
+
| IK34:pccB_fw || 2013-07-19 || Forward primer for pccB || ATGTCCGAGCCGGAAGAGC
|-
|-
-
| PW37:pSB1C3-TycC5dC_rev || Sigma-Aldrich Chemie GmbH || VC00021`21841361
+
| IK35:BBa_J04450-pccB_rev || 2013-07-19 || Reverse primer for mRFP-containing backbones with Gibson overhang for pccB || ATGTCGGGCTGCTGCTCTTCCGGCTCGGACAT CTAGTATTTCTCCTCTTTCTCTAG
|-
|-
-
| AT101 || Sigma-Aldrich Chemie GmbH  || VC00021`21865217
+
| IK36:pLF03_seq_rev || 2013-07-26 || reverse primer for pLF03 for sequencing || CCGGTATCAACAGGGACACCAG
|-
|-
-
| AT102 || Sigma-Aldrich Chemie GmbH || VC00021`21865218
+
| IK37:pLF03-catR_seq_rev || 2013-07-26 || reverse primer for pLF03 sequencing || CATTGAGCAACTGACTGAAATGCCTC
|-
|-
-
| AT103 || Sigma-Aldrich Chemie GmbH || VC00021`21865219
+
| IK38:pIK1_fw || 2013-08-30 || Forward mutagenic primer for pIK1 || ATTCCCTGGCCTTGT t GGTCAATCTGGGGATTTATG
|-
|-
-
| AT104 || Sigma-Aldrich Chemie GmbH || VC00021`21865220
+
| IK39:pIK_rev || 2013-08-30 || Reverse mutagenic primer for pIK1 || CCCAGATTGACC a ACAAGGCCAGGGAATGAATCTTC
|-
|-
-
| AT105 || Sigma-Aldrich Chemie GmbH || VC00021`21865221
+
| IK40:pIK2-BBa_J23114-BBa_B0030_rev || 2013-09-06 || reverse primer for pIK2 with Gibson overhang for BBa_J23114-BBa_B0030 || TTTAATCTCTAGAGCTAGCATTGTACCTAGGACTGAGCTAGCCATAAA CTCTAGTAGAGAGCGTTCAC
|-
|-
-
| AT106 || Sigma-Aldrich Chemie GmbH || VC00021`21865222
+
| IK41:BBa_I746200-BBa_B0030-BBa_J23114_fw || 2013-09-06 || forward primer for BBa_I746200 with Gibson overhang for BBa_B0030-BBa_J23114 || ATGCTAGCTCTAGAGATTAAAGAGGAGAAATACTAG ATGAACAAGAAGATTCATTCCCTG
|-
|-
-
| AT107 || Sigma-Aldrich Chemie GmbH || VC00021`21865223
+
| IK42:BBa_I746200_rev || 2013-09-06 || reverse primer for BBa_I746200 || TCAGAAGTGGGTGTTTACGCTC
|-
|-
-
| AT108 || Sigma-Aldrich Chemie GmbH || VC00021`21865224
+
| IK43:pIK2-BBa_I746200_fw || 2013-09-06 || forward primer for pIK2 with Gibson overhang for BBa_I746200 || TGAGCGTAAACACCCACTTCTGA TACTAGAGTCACACTGGCTC
|-
|-
-
| IK40:pIK2-BBa_J23114 || Sigma-Aldrich Chemie GmbH || VC00021`21871657
+
|NK_01_FS_62_screening_L_BB(without_mRFP)_rv||2013-08-02 ||Primer for screening/sequencing of pFSN construct||GTTCACCGACAAACAACAGATAAAACG
|-
|-
-
| IK41:BBa_I746200-Bba || Sigma-Aldrich Chemie GmbH || VC00021`21871658
+
| ygfG21C1 || 2013-06-03 || For insertion of methylmalonyl-CoA synthesis pathway into ''E. coli''. Primer from <bib id="pmid17959404"/>. || TCACCGCGCACCGGCCTGCGGCAGCTCAGTGAATTTATCC AGATCTCGATCCC
|-
|-
-
| IK42:BBa_I746200_rev || Sigma-Aldrich Chemie GmbH || VC00021`21871659
+
| ygfG21C2 || 2013-06-03 || For insertion of methylmalonyl-CoA synthesis pathway into ''E. coli''. Primer from <bib id="pmid17959404"/>. || GTTATGCTGGATAATTTCTGCCGCTTCATTGGCGGTCATCC CAAAAAACCCCTCAAG
|-
|-
-
| IK43:pIK2-BBa_I746200_fw || Sigma-Aldrich Chemie GmbH || VC00021`21871660
+
|}
 +
 
 +
==Module shuffling==
 +
 
 +
{|class="wikitable"
|-
|-
-
| HM20_BB-pSB6A1_ || Eurofins MWG GmbH || 27080132
+
! Identifier !! Order date !! Note !! Sequence
|-
|-
-
| FS_77_rv || Eurofins MWG GmbH || 27080134
+
| AT01:RFC10prefix_TycA_fw|| 2013-08-12 || Fw primer for amplification of TycAdCom; introduction of RFC10 prefix || TTTT GAATTC GCGGCCGC T TCTAG ATG TTA GCA AAT CAa GCC AAT C
|-
|-
-
| NI19:ngrA-BB_fw || Eurofins MWG GmbH || 27114029
+
| AT02:RFC10suffix_TycA_rv
 +
|| 2013-08-12 || Rv primer for amplification of TycAdCom; introduction of RFC10 suffix|| TTTT CTGCAG CGGCCGC T ACTAGT A aGT TCG tTC TAC TTC TTT TTT C
|-
|-
-
| NI20:ngrA_rv || Eurofins MWG GmbH || 27114030
+
| AT03:RFC10prefix-TycB1_fw
 +
|| 2013-08-12 || Fw primer for amplification of TycB1dCom; introduction of RFC10 prefix|| TTTT GAATTC GCGGCCGC T TCTAG ATG AGT GTA TTT AGC AAA GAA CAA G
|-
|-
-
| NI21:ngrA_fw || Eurofins MWG GmbH || 27114031
+
| AT04:RFC10suffix_TycB1_rv
 +
|| 2013-08-12 || Rv primer for amplification of TycB1dCom; introduction of RFC10 suffix || TTTT CTGCAG CGGCCGC T ACTAGT A TTC CTC CCC aCC TTC
|-
|-
-
| NI22:ngrA_rv || Eurofins MWG GmbH || 27114032
+
| AT05:RFC10prefix-TycC5_fw
 +
|| 2013-08-12 || Fw primer for amplification of TycC5; introduction of RFC10 prefix || TTTT GAATTC GCGGCCGC T TCTAG AG GCG CAT ATT GCa GAG AG
|-
|-
-
| VF2 || Eurofins MWG GmbH || 27114033
+
| AT06:RFC10suffix_TycC5_rv
 +
|| 2013-08-14 || Rv primer for amplification of TycC5; introduction of RFC10 suffix || TTTT CTGCAG CGGCCGC T ACTAGT A TTT GGC TGT CTC TTC GAT GAA C
|-
|-
-
| HM_11:lacI_RBS(1)_DelH_rev || Sigma-Aldrich Chemie GmbH || VC00021`22003550
+
| AT07:RFC10prefix-TycC6_fw
 +
|| 2013-08-12 || Fw primer for amplification of TycC6; introduction of RFC10 prefix  || TTTT GAATTC GCGGCCGC T TCTAG AG GGG AAT GTC TTC TCG ATC
|-
|-
-
| AR07:TycC4dCom_short-BB_rev || Sigma-Aldrich Chemie GmbH || VC00021`22008078
+
| AT08:RFC10suffix_TycC6_rv
 +
|| 2013-08-14 || Rv primer for amplification of TycC6; introduction of RFC10 suffix || TTTT CTGCAG CGGCCGC T ACTAGT A TTA TTT CAG GAT aAA CAG TTC TTG
|-
|-
-
| AR08:TycC4dCom_long-BB_rev || Sigma-Aldrich Chemie GmbH || VC00021`22008079
+
| AT09:R10_B1_fw_longer
 +
|| 2013-08-18 || Fw primer (longer) for amplification of TycB1dCom; introduction of RFC10 prefix  || TTTT GAA TTC GCG GCC GCT TCT AG    ATG AGT GTA TTT AGC AAA GAA CAA GTT C
|-
|-
-
| AR02:c(TycC2)s_rev || Sigma-Aldrich Chemie GmbH || VC00021`22008073
+
| AT10:R10_B1_rv_longer
 +
|| 2013-08-18 || Rv primer (longer for amplification of TycB1dCom; introduction of RFC10 suffix || TTTT CTG CAG CGG CCG CTA CTA GTA ATA CGC aCT TTC CTC CCC GCC
|-
|-
-
| AR04:c(TycC2)l_rev || Sigma-Aldrich Chemie GmbH || VC00021`22008075
+
| AT11:R10_C5_fw_rpos
 +
|| 2013-08-18 || Fw primer (repositioned) for amplification of TycC6; introduction of RFC10 prefix|| TTTT GAATTC GCGGCCGC T TCTAG AG GAGCAGTTCGAGACGATCCAGCC
|-
|-
-
| AR05::l-Ind_fw || Sigma-Aldrich Chemie GmbH || VC00021`22008076
+
| AT101|| 2013-09-05|| forward screening primer TycAdCom || GGACATCATCGAACAGGC
|-
|-
-
| AR03:s-Ind_fw || Sigma-Aldrich Chemie GmbH || VC00021`22008074
+
| AT102 || 2013-09-05|| reverse screening primer TycAdCom || GACATAGGCAAGATCCGTAG
|-
|-
-
| AR01:sTycC4dC_fw || Sigma-Aldrich Chemie GmbH || VC00021`22008072
+
| AT103|| 2013-09-05|| forward screening primer TycAdCom || GCAAGGACAAGTAGATGGC
|-
|-
-
| AR06:lTycC4dC_fw || Sigma-Aldrich Chemie GmbH || VC00021`22008077
+
| AT104 || 2013-09-05|| reverse screening primer TycAdCom || GTACAGCCATACAGTCGC
|-
|-
-
| RB92:entF-T3_fw || Sigma-Aldrich Chemie GmbH || VC00021`22020305
+
| AT105 || 2013-09-05|| forward screening primer TycC5 || GTCACCATGCTGAGAACG
|-
|-
-
| RB93:tycA1-T3_fw || Sigma-Aldrich Chemie GmbH || VC00021`22020306
+
| AT106 || 2013-09-05|| reverse screening primer TycC5 || CAGCAAAGCGGTCATAATC
|-
|-
-
| RB93:tycC6-T3_fw || Sigma-Aldrich Chemie GmbH || VC00021`22020307
+
| AT107 || 2013-09-05|| forward screening primer TycC6 || AAGCTACGCTGTTGATTGC
|-
|-
-
| RB94:delH4-T3_fw || Sigma-Aldrich Chemie GmbH || VC00021`22020308
+
| AT108 || 2013-09-05|| reverse screening primer TycC6 || AGCACGTAACGATCCTC
|-
|-
-
| RB95:delH5-T3_fw || Sigma-Aldrich Chemie GmbH || VC00021`22020309 
+
| IK09:TycA_A1_fw || 2013-07-08 || Colony-PCR of ''Brevibacillus parabrevis'': TycA A domain || ATGTTAGCAAATCAAGCCAATCTC
|-
|-
-
| RB96:plu2642-T3_fw || Sigma-Aldrich Chemie GmbH || VC00021`22020310
+
| IK10:TycA_A1_rev || 2013-07-08 || Colony-PCR of ''Brevibacillus parabrevis'': TycA A domain || TTGGTTTGCTGTAAGATCAGGCTC
|-
|-
-
| RB97:plu2670-T3_fw || Sigma-Aldrich Chemie GmbH || VC00021`22020311
+
| IK11:TycB_A1_fw || 2013-07-08 || Colony-PCR of ''Brevibacillus parabrevis'': TycB A1 domain || AATTCGGGAGTCGAGCTTTGTCAG
|-
|-
-
| RB107:tycC2_A_fw || Sigma-Aldrich Chemie GmbH || VC00021`22098771
+
| IK12:TycC6_rev || 2013-07-19 || reverse primer for tyrocidine TycC6 module || TTATTTCAGGATGAACAGTTCTTGCAGG
|-
|-
-
| RB106:tycC2_T3_rv || Sigma-Aldrich Chemie GmbH || VC00021`22098770
+
| IK13:IK13:TycB1-dCom-dC_fw || 2013-07-19 || forward primer for tyrocidine TycB1 module without Com and C domain || GATTGCGTGGCAAACAATTCGGGAGTC
|-
|-
-
| RB99:plu2642_pRB23-plu_fw || Sigma-Aldrich Chemie GmbH || VC00021`22098763
+
| IK14:TycB1-TycC6_rev || 2013-07-19 || reverse primer for tyrocidine TycB1 module with Gibson overhang for TycC6 || CAGGCTCGATCGAGAAGACATTCCC TTCCTCCCCGCCTTCCACATACGC
|-
|-
-
| RB100:plu2642_pRB23-plu_rv || Sigma-Aldrich Chemie GmbH || VC00021`22098764
+
| IK15:TycC6-TycB1_fw || 2013-07-19 || forward primer for tyrocidine TycC6 module with Gibson overhang for TycB1 || GCGTATGTGGAAGGAGGGGAGGAA GGGAATGTCTTCTCGATCGAGCCTG
|-
|-
-
| RB103:plu2642_valInd_fw || Sigma-Aldrich Chemie GmbH || VC00021`22098767
+
| IK16:TycA_fw || 2013-07-19 || forward primer for tyrocidine TycA module || ATGTTAGCAAATCAGGCCAATCTCATC
|-
|-
-
| RB101:plu2642_A_rv || Sigma-Aldrich Chemie GmbH || VC00021`22098765
+
| IK17:TycA-dCom-TycC5_rev || 2013-07-19 || reverse primer for tyrocidine TycA module without Com domain with Gibson overhang for TycC5 || GAATGCGCTCTCGGCAATATGGGC TGTTCGCTCTACTTCTTTTTTCTCGG
|-
|-
-
| RB105:tycC2_T3_fw || Sigma-Aldrich Chemie GmbH || VC00021`22098769
+
| IK18:TycC5-TycA-dCom_fw || 2013-07-19 || forward primer for tyrocidine TycC5 module with Gibson overhang for TycAdCom || CCGAGAAAAAAGAAGTAGAGCGAACA GCCCATATTGCCGAGAGCGCATTC
|-
|-
-
| RB104:plu2642_valInd_rv || Sigma-Aldrich Chemie GmbH || VC00021`22098768
+
| IK19:TycB1-TycC5_rev || 2013-07-19 || reverse primer for tyrocidine TycB1 module with Gibson overhang for TycC5 || GAATGCGCTCTCGGCAATATGGGC TTCCTCCCCGCCTTCCACATACGC
|-
|-
-
| RB102:indC_Ox_fw || Sigma-Aldrich Chemie GmbH || VC00021`22098766
+
| IK20:TycC5-TycB1_fw || 2013-07-19 || forward primer for tyrocidine TycC5 module with Gibson overhang for TycB1 || GCGTATGTGGAAGGAGGGGAGGAA GCCCATATTGCCGAGAGCGCATTC
|-
|-
-
| RB108:tycC2_A_rv || Sigma-Aldrich Chemie GmbH || VC00021`22098772
+
| IK21:pSB4K5-TycB1dComdC_rev || 2013-07-19 || reverse primer for mRFP-carrying backbones with Gibson overhang for tyrocidine TycA module (nomenclature wrong) || GTCGATGAGATTGGCCTGATTTGCTAACAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
|-
|-
-
| RB109:tycC2_valInd_fw || Sigma-Aldrich Chemie GmbH || VC00021`22098773
+
| IK22::pSB4K5-TycC6_fw || 2013-07-19 || forward primer for mRFP-carrying backbones with Gibson overhang for tyrocidine TycC6 module || AACATCCTGCAAGAACTGTTCATCCTGAAA TAATAACGCTGATAGTGCTAGTGTAGATC
|-
|-
-
| RB110:plu2642_TE_fw || Sigma-Aldrich Chemie GmbH || VC00021`22098774
+
| IK23:pSB4K5-TycA_rev || 2013-07-19 || reverse primer for mRFP-carrying backbones with Gibson overhang for tyrocidine TycB1 module without Com and C domains + start codon (nomenklature wrong) || GACTCCCGAATTGTTTGCCACGCAATCCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
|-
|-
-
| indC-BLAST || IDT || 65647038
+
| PW01:TycB_A1_rev || 2013-07-08 || Colony-PCR of ''Brevibacillus parabrevis'': TycB A1 domain || CTTGGCACTTCCTTCAGGCTTC
|-
|-
-
| indC-T-Plum-BLAST || IDT || 65647039
+
| PW02:TycB_E1_fw || 2013-07-08 || Colony-PCR of ''Brevibacillus parabrevis'': TycB E domain || CGAGAGAGCGAGCAAGGTG
|-
|-
-
| indC-T-BLAST || IDT || 65647040
+
| PW03:TycB_E1_rev || 2013-07-08 || Colony-PCR of ''Brevibacillus parabrevis'': TycB E domain || GTACTCGCCTTCTTCTTTTGC
|-
|-
-
| project5-BLAST || IDT || 65647041
+
| PW04:pSB1C3-TycC5ΔC_rev || 2013-07-19 || Integration of Tetrapeptide NRPS from ''Brevibacillus parabrevis'' in backbone; Gibson primer for Tetrapeptide I & II || TGTTTTGGTTGCGAGGAAGCTGTGCAGCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
|-
|-
-
| random1_indC || IDT || 65647042
+
| PW05:TycC5ΔC_fwd || 2013-07-19 || Amplification of TycC5-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide I & II || ATGCTGCACAGCTTCCTCGCAACCAAAACAGCC
|-
|-
-
| random2_project5-BLAST || IDT || 65647043
+
| PW06:TycC5ΔC-TycB1ΔCom_rev || 2013-07-19 || Amplification of TycC5-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide I & II || GTGAAACAGCATCCCCTCTTGCATCGG AGGCTCGATCGAGAAGACATTCCCTTTG
|-
|-
-
| random_indC-BLAST || IDT || 65647044
+
| PW07:TycC5ΔC-TycB1ΔCom_fwd || 2013-07-19 || Amplification of TycB1+C(TycB2)-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide I & II || CAAAGGGAATGTCTTCTCGATCGAGCCT CCGATGCAAGAGGGGATGCTGTTTCAC
|-
|-
-
| RB90:indC-T-screen_fw || IDT || 65775415
+
| PW08:C(TycB2)-TycAΔCom_rev || 2013-07-19 || Amplification of TycB1+C(TycB2)-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide I & II || GTTGTCGATGAGATTGGCCTGATTTGCTAACAT GATTTGCGCCAGCTCCTGCTCCGTGTT
|-
|-
-
| RB91:bpsA-T-screen_fw || IDT || 65775416
+
| PW09:C(TycB2)-TycAΔCom_fwd || 2013-07-19 || Amplification of TycA-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide I & II || AACACGGAGCAGGAGCTGGCGCAAATC ATGTTAGCAAATCAGGCCAATCTCATCGACAAC
|-
|-
-
| PW38:CCDB_fwd || IDT || 65775417
+
| PW10:TycAΔCom-TycC6_rev || 2013-07-19 || Amplification of TycAΔCom-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide I || CTTTGGCTGTCTCTTCGATGAACGC TCGCTCTACTTCTTTTTTCTCGGTGCAATG
|-
|-
-
| PW39:pSB1C3-CCDB_rev || IDT || 65775418
+
| PW11:TycAΔCom-TycC6_fwd || 2013-07-19 || Amplification of TycC6-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide I || CATTGCACCGAGAAAAAAGAAGTAGAGCGA GCGTTCATCGAAGAGACAGCCAAAG
|-
|-
-
| PW40:CCDB-C(TycC2)_fwd || IDT || 65775419
+
| PW12:TycAΔE-TycC6_rev || 2013-07-19 || Amplification of TycAΔE-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide II; one mismatch C->t || CTTTTGCACAGGCTCGATCGAGAAGAC GCTtTTGACAAAAAGAGCAACCTG
|-
|-
-
| PW41:CCDB-C(TycC2)_rev || IDT || 65775420
+
| PW13:TycAΔE-TycC6_fwd || 2013-07-19 || Amplification of TycC6-Module from ''Brevibacillus parabrevis''; Gibson primer for Tetrapeptide II; one mismatch G->a || CAGGTTGCTCTTTTTGTCAAaAGC GTCTTCTCGATCGAGCCTGTGCAAAAG
|-
|-
-
| HM21_fw_lacI_BbsI_Xba || IDT || 65775426
+
| PW14:C(TycC2)-indC_rev || 2013-08-16 || Amplification of C-domain from TycC2 from ''Brevibacillus parabrevis''; Gibson overhang to IndC; for construct 1, 2 & 3 || ACATTGTGTAATATTATTTTCTAACAT CGTTTTGCTGCTGGCAGGCTG
|-
|-
-
| HM22_rev_RBS || IDT || 65775427
+
| PW15:C(TycC2)-indC_fwd || 2013-08-16 || Amplification of indC from ''Photorhabdus luminescens''; Gibson overhang to C-domain from TycC2; for construct 1, 2 & 3 || CAGCCTGCCAGCAGCAAAACG ATGTTAGAAAATAATATTACACAATGT
|-
|-
-
| HM23_fw_RBS || IDT || 65775428
+
| PW16:indC_rev || 2013-08-16 || Amplification of indC from ''Photorhabdus luminescens''; no Gibson overhang; for construct 1, 2 & 3 || TTAGATTATTTTCTCAATCTCAGCAACACCTTC
|-
|-
-
| HM24_rev_BbsI_MfeI || IDT || 65775429
+
| PW17:TycAdE-C(TycC2)_rev || 2013-08-16 || Amplification of TycAdE from ''Brevibacillus parabrevis''; Gibson overhang to C-domain from TycC2; for construct 1 || CGAAAGGAAGCGGGCCAGCTC AGCAACCTGCTCGATCGTCGGGTA
|-
|-
-
| PW42:pSB1C3-CCDB_rev_shorter || IDT || 65775430
+
| PW18:TycAdE-C(TycC2)_fwd || 2013-08-16 || Amplification of C-domain from TycC2-module from ''Brevibacillus parabrevis''; Gibson overhang to TycAdE; for construct 1 || TACCCGACGATCGAGCAGGTTGCT GAGCTGGCCCGCTTCCTTTCG
|-
|-
-
| delH_f1_PacI_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW19:TycC4dC_fwd || 2013-08-16 || Amplification of TycC4-module from ''Brevibacillus parabrevis'' without the C-domain; no Gibson overhang, ATG added; for construct 3 || ATGTATCCGCGCGATCTGACGATTC
|-
|-
-
| delH_f1_SalI_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW20:TycC4dC-C(TycC2)_rev || 2013-08-16 || Amplification of TycC4-module from ''Brevibacillus parabrevis'' without the C-domain; Gibson overhang to C-domain form TycC2; for construct 3 || GGTGTACTCGGTTTTTTCCGA AATATGCGCAGCCAACTCATG
|-
|-
-
| delH_f2_SalI_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW21:TycC4dC-C(TycC2)_fwd || 2013-08-16 || Amplification of C-domain from TycC2-module from ''Brevibacillus parabrevis''; Gibson overhang to TycC4dC; for construct 3 || CATGAGTTGGCTGCGCATATT TCGGAAAAAACCGAGTACACC
|-
|-
-
| delH_f2_KpnI_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW22:pSB1C3-TycC4dC_rev || 2013-08-16 || Insertion of construct 3 into pSB1C3-backbone, amplification of pSB1C3; Gibson overhang to TycC4dC; for construct 3 || GAATCGTCAGATCGCGCGGATACAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
|-
|-
-
| AraCbb_KpnI_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW23:indC-pSB1C3_fwd || 2013-08-16 || Insertion of either fragments into pSB1C3-backbone, amplification of pSB1C3; Gibson overhang to indC from ''Photorhabdus luminescens''; for constructs 1, 2 & 3 || GGTGTTGCTGAGATTGAGAAAATAATCTAA TAATAACGCTGATAGTGCTAGTGTAGATC
-
|-  
+
-
| AraCbb_PacI_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
|-
|-
-
| Screen_delH_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW24:TycC1dC_fwd || 2013-08-16 || Amplification of the TycC1-module from ''Brevibacillus parabrevis'' without the C-domain; no Gibson overhang, ATG added; for construct 2 || ATGCAGACGAACAAACAACAGACG
|-
|-
-
| Screen_delH_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW25:pSB1C3-TycC1dC || 2013-08-16 || Insertion of construct 2 in pSB1C3-backbone, amplification of pSB1C3; Gibson overhang to TycC1-module without C-domain; for construct 2 || CGTCTGTTGTTTGTTCGTCTGCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
|-
|-
-
| AraCbb_PacI_rev2 || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW26:TycC5dC_fwd || 2013-09-04 || Amplification of TycC5-module without C-domain from ''Brevibacillus parabrevis''; no Gibson overhang, ATG added; for constructs A & B || ATGCTGCACAGCTTCCTCGCAACC
|-
|-
-
| DelH_EcoRI_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW27:TycC5dC-C(TycC4)_rev || 2013-09-04 || Amplification of TycC5-module without C-domain from ''Brevibacillus parabrevis''; Gibson overhang to C-domain from TycC4; for constructs A & B || CACATACGTCTCTTTTCCGCTCGT TTCGATGAACGCCGCCAGTTC
|-
|-
-
| DelH_EcoRI_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW28:TycC5dC-C(TycC4)_fwd || 2013-09-04 || Amplification of C-domain from TycC4-module from ''Brevibacillus parabrevis''; Gibson overhang to TycC5-module without C-domain; for constructs A & B || GAACTGGCGGCGTTCATCGAA ACGAGCGGAAAAGAGACGTATGTG
|-
|-
-
| ygfG21C1 || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW29:C(TycC4)-TycC4dC_rev || 2013-09-04 || Amplification of C-domain from TycC4-module from ''Brevibacillus parabrevis''; Gibson overhang to TycC4-module without C-domain; for constructs A, B, C, E & G || GAATCGTCAGATCGCGCGGATA GGCAAACGTGTTGTTGAAATC
|-
|-
-
| ygfG21C2 || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW30:C(TycC4)-TycC4dC_fwd || 2013-09-04 || Amplification of TycC4-module without C-domain from ''Brevibacillus parabrevis''; Gibson overhang to C-domain from TycC4-module; for constructs A, B, C, E & G || GATTTCAACAACACGTTTGCC TATCCGCGCGATCTGACGATTC
|-
|-
-
| delH_f1_fw_long || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW31:TycC4-C(TycC4)_rev || 2013-09-04 || Amplification of TycC4-module from ''Brevibacillus parabrevis''; Gibson overhang to C-domain from TycC4-module; for constructs B & C || CACATACGTCTCTTTTCCGCTCGT GGCAATATGCGCAGCCAACTCATG
|-
|-
-
| delH_f1_fw_short || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW32:TycC4-C(TycC4)_fwd || 2013-09-04 || Amplification of C-domain from TycC4-module from ''Brevibacillus parabrevis''; Gibson overhang to TycC4-module; for constructs B & C || CATGAGTTGGCTGCGCATATTGCC ACGAGCGGAAAAGAGACGTATGTG
|-
|-
-
| delH_f1_PacI_fw_short || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW33:TycC6dTE-C(TycC2)_rev || 2013-09-04 || Amplification of TycC6-module without the TE-domain from ''Brevibacillus parabrevis''; Gibson overhang to C-domain from TycC2-module; for constructs D & F || GGTGTACTCGGTTTTTTCCGA CGTGATGAAATCGGCCACCTTTTC
|-
|-
-
| delH_f1_fw_short2 || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW34:TycC6dTE-C(TycC2)_fwd || 2013-09-04 || Amplification of C-domain from TycC2-module from ''Brevibacillus parabrevis''; Gibson overhang to TycC6-module without TE-domain; for constructs D & F || GAAAAGGTGGCCGATTTCATCACG TCGGAAAAAACCGAGTACACC
|-
|-
-
| IK01:pLF03_integr_argK_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW35:TycC6dTE-C(TycC4)_rev || 2013-09-04 || Amplification of TycC6-module without the TE-domain from ''Brevibacillus parabrevis''; Gibson overhang to C-domain from TycC4-module; for constructs E & G || CACATACGTCTCTTTTCCGCTCGT CGTGATGAAATCGGCCACCTTTTC
|-
|-
-
| IK02:pLF03_integr_pET21c_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW36:TycC6dTE-C(TycC4)_fwd || 2013-09-04 || Amplification of C-domain from TycC4-module from ''Brevibacillus parabrevis''; Gibson overhang to TycC6-module without TE-domain; for constructs E & G || GAAAAGGTGGCCGATTTCATCACG ACGAGCGGAAAAGAGACGTATGTG
|-
|-
-
| IK03:pLF03_integr_argK_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW37:pSB1C3-TycC5dC_rev || 2013-09-04 || Insertion of constructs A & B in pSB1C3-backbone, amplification of pSB1C3; Gibson overhang to TycC5-module without C-domain; for constructs A & B || GGTTGCGAGGAAGCTGTGCAGCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
 +
|}
 +
 
 +
==Linker Variation==
 +
{|class="wikitable"
|-
|-
-
| IK04:pLF03_integr_argK_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
! Identifier !! Order date !! Note !! Sequence
|-
|-
-
| IK05:pLF03_integr_ygfG_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
|AR01|| 2013-09-17 || sTycC4dC_fw || ATG AAGCACTTGCTGCCGCTCGTC
|-
|-
-
| IK06:pLF03_integr_ygfG_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
|AR02|| 2013-09-17 ||c(TycC2)s_rev|| TTGTGTAATATTATTTTCTAACAT CGCTACCTGTTGCAACCGCTC
|-
|-
-
| IK07:ygfG21C1 || Sigma-Aldrich Chemie GmbH || VC00021
+
|AR03|| 2013-09-17 ||s-Ind_fw|| GAGCGGTTGCAACAGGTAGCG ATGTTAGAAAATAATATTACAATGT
|-
|-
-
| IK08:ygfG21C2 || Sigma-Aldrich Chemie GmbH || VC00021
+
|AR04|| 2013-09-17 ||c(TycC2)l_rev_l||TTGTGTAATATTATTTTCTAACAT AAGCGACTGCTGGTTCGCG
|-
|-
-
| FS_01_pSB4K5_DelA_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|AR05|| 2013-09-17 ||l-Ind_fw||AACGCGAACCAGCAGTCGCTT ATGTTAGAAAATAATATTACACAATGT
|-
|-
-
| FS_11_DelAG_5_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|AR06|| 2013-09-17 ||lTycC4dC_fw||ATG CGCAACTATCCGGTCGAGACG
|-
|-
-
| FS_13_DelOP_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|AR07|| 2013-09-17 ||TycC4dCom_short-BB_rev||GACGAGCGGCAGCAAGTGCTTCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
|-
|-
-
| FS_15_DelL_mRFP_pSB4K5_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|AR08|| 2013-09-17 ||TycC4dCom_long-BB_rev||CGTCTCGACCGGATAGTTGCGCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
|-
|-
-
| FS_02:DelAG1_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW19:TycC4dC_fwd || 2013-08-16 || Amplification of TycC4-module from ''Brevibacillus parabrevis'' without the C-domain; no Gibson overhang, ATG added; for construct 3 || ATGTATCCGCGCGATCTGACGATTC
|-
|-
-
| FS_03:DelAG1_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW20:TycC4dC-C(TycC2)_rev || 2013-08-16 || Amplification of TycC4-module from ''Brevibacillus parabrevis'' without the C-domain; Gibson overhang to C-domain form TycC2; for construct 3 || GGTGTACTCGGTTTTTTCCGA AATATGCGCAGCCAACTCATG
|-
|-
-
| FS_04:DelAG2_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW21:TycC4dC-C(TycC2)_fwd || 2013-08-16 || Amplification of C-domain from TycC2-module from ''Brevibacillus parabrevis''; Gibson overhang to TycC4dC; for construct 3 || CATGAGTTGGCTGCGCATATT TCGGAAAAAACCGAGTACACC
|-
|-
-
| FS_05:DelAG2_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
| PW14:C(TycC2)-indC_rev || 2013-08-16 || Amplification of C-domain from TycC2 from ''Brevibacillus parabrevis''; Gibson overhang to IndC; for construct 1, 2 & 3 || ACATTGTGTAATATTATTTTCTAACAT CGTTTTGCTGCTGGCAGGCTG
|-
|-
-
| FS_06:DelAG_3_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
|}
-
|-
+
 
-
| FS_07:DelAG_3_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
==Domain shuffling and PPTases==
-
|-
+
 
-
| FS_08:DelAG_4_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
{|class="wikitable"
-
|-
+
-
| FS_09:DelAG_4_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_10:DelAG_5_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_12:DelOP_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_14:DelL_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_16:mRFP_pSB4K5_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB05:RBS1-BpsA_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB06:BpsA-RBS2_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB07:RBS2-svp_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB08:svp-pSB1C3_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB09:pSB1C3_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB10:pSB1C3-RBS1_rv  || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_11: DelAG_5_short_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_13: DelL_short_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_15: DelOP_short_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK_09:TycA_A1_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK_10:TycA_A1_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK_19:TycB_A1_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW_04:TycB_A1_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW_02:TycB_E1_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW_03:TycB_E1_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB11:BpsA_Taka_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB12:BpsA_Taka_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB13:svp_Taka_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB14:svp_Taka_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB15:svp_Sanch_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB16:svp_Sanch_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB17:sfp_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB18:sfp_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB19:entD_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB20:entD_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM02_DelH_Gib1.1_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM03_DelH_Gib1.2_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM04_delH_Gib1.2_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM05_DelH_Gib2.1_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM06_DelH_Gib2.1_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM07_DelH_Gib2.2_fw  || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM08_DelH_Gib2_end_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM09_AraC_RBS_Delh_rev  || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| HM10_RBS_lacZ_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_20: DelF_fw  || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_21: DelF_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_22: DelOP_short || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_24: DelAE || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_25: DelEF || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_26: DelFG_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK24:pccB_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK25:pccB_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| NI11:BpsA_rvN || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| NI12:svp_fwN || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB21:pSB1C3_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB22:pSB1C3_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB27:indC-Plum_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB28:indC-Plum_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB35:sfp-Naka_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB36:sfp-Naka_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB37:Plum-extr_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB38:Plum-extr_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB43:Bsub-extr_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| RB44:Bsub-extr_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| FS_23: DelG_OP_long_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK12:TycC6_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK13:TycB1-dCom-dC_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK14:TycB1-TycC6_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK15:TycC6-TycB1_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK16:TycA_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK17:TycA-dCom-TycC5_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK18:TycC5-TycA-dCom_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK19:TycB1-TycC5_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK20:TycC5-TycB1_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK21:pSB4K5-TycB1dComdC_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK22:pSB4K5-TycC6_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| IK23:pSB4K5-TycA_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW04:pSB4K5-TycC5dC_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW05:TycC5dC_fwd || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW06:TycC5dC-TycB1dCom_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW07:TycC5dC-TycB1dCom_fwd || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW08:C(TycB2)-TycAdCom_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW09:C(TycB2)-TycAdCom_fwd || Sigma-Aldrich Chemie GmbH || VC00021
+
-
|-
+
-
| PW10:TycAdCom-TycC6_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
|-
|-
-
| PW11:TycAdCom-TycC6_fwd || Sigma-Aldrich Chemie GmbH || VC00021
+
! Identifier !! Order date !! Note !! Sequence
|-
|-
-
| PW12:TycAdE-TycC6_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| KH1_ccdb_fw || 2013-07-23 || amplifing ccdb with RBS from pDONR || TGGTGCCATTACATACAGATACT GAGAACAGGGGCTGGTGAAATGC
 +
|-
 +
| KH2_ccdb_rv || 2013-07-23 || amplifing ccdb with RBS from pDONR || AGAGTCTGTCTGTTCAATCCACTT TTATATTCCCCAGAACATCAGGTTAATGGCG
 +
|-
 +
| KH3_indC_backbone-withoutT_fw || 2013-07-23 || linearization of vector with partial indigoidine synthase || AAGTGGATTGAACAGACAGACTCTAAAAC
 +
|-
 +
| KH4_indC_backbone-withoutT_rv || 2013-07-23 || linearization of vector with partial indigoidine synthase || AGTATCTGTATGTAATGGCACCAATAGACGC
 +
|-
 +
| KH5_indC_T_fw || 2013-07-23 || introducing T domain of indC || TGGTGCCATTACATACAGATACT GAAATAAGGCTTGGAAAAATTTGGATGGAAGT
 +
|-
 +
| KH6_indC_T_rv || 2013-07-23 || introducing T domain of indC || AGAGTCTGTCTGTTCAATCCACTT AGCCAATTCTGCTATATTAGGAGATTGA
 +
|-
 +
| KH7_bpsA_T_fw || 2013-07-23 || introducing T domain of bpsA || TGGTGCCATTACATACAGATACT GAGAAGGAAATCGCAGCCGTGTGGG
 +
|-
 +
| KH8_bpsA_T_rv || 2013-07-23 || introducing T domain of bpsA || AGAGTCTGTCTGTTCAATCCACTT GGCCAGCTTTTCAATTGTTGGTGACTCC
 +
|-
 +
| KH9_ccdB-Big_fw || 2013-08-06 || ccdB_Big amplified from pDONOR (Dominik) for indC(ccdB) construct || TGCCATTACATACAGATACT ACTGGCTGTGTATAAGGGAGCCTGAC
 +
|-
 +
| KH10_ccdB-Big_rv || 2013-08-06 || ccdB_Big amplified from pDONOR (Dominik) for indC(ccdB) construct || AGAGTCTGTCTGTTCAATCCACTT CGCGTGGATCCGGCTTAC
 +
|-
 +
| KH11_plu2642-T_fw || 2013-08-14 || T domain exchange plu2642 || TGGTGCCATTACATACAGATACT GAAACACAGATCGTAAAGATATGG
 +
|-
 +
| KH12_plu2642-T_rv || 2013-08-14 || T domain exchange plu2642 || AGAGTCTGTCTGTTCAATCCACTT TGCCAATTGTTTAACCGTTG
 +
|-
 +
| KH13_BBa-entD_fw || 2013-08-09 || submitting endD to registry || GCAT GAATTCGCGGCCGCTTCTAG ATGAAAACTACGCATACCTCCCTC
 +
|-
 +
| KH14_BBa-entD_rv || 2013-08-09 || submitting endD to registry || GCAT CTGCAGCGGCCGCTACTAGTA TCATTAATCGTGTTGGCACAGCG
 +
|-
 +
| KH15_BBa-sfp_Bsub_fw || 2013-08-09 || submitting sfp_Bsub to registry || GCAT GAATTCGCGGCCGCTTCTAG ATGAAGATTTACGGAATTTATATGGAC
 +
|-
 +
| KH16_BBa-sfp_Bsub_rv || 2013-08-09 || submitting sfp_Bsub to registry || GCAT CTGCAGCGGCCGCTACTAGTA TCATTATAAAAGCTCTTCGTACGAGAC
 +
|-
 +
| KH17_BBa-svp_Svert_fw || 2013-08-09 || submitting svp_Svert to registry '''needed template: pRB04''' || GCAT GAATTCGCGGCCGCTTCTAG A TGATCGCCGCCCTCCTGCCCTCC
 +
|-
 +
| KH18_BBa-svp_Svert_rv || 2013-08-09 || submitting svp_Svert to registry '''needed template: pRB04''' || GCAT CTGCAGCGGCCGCTACTAGTA AGCTAGCTCATTACGGGAC
 +
|-
 +
| KH19_BBa-delC_fw || 2013-08-09 || submitting delC to registry || GCAT GAATTCGCGGCCGCTTCTAG ATGCAGCTCGTGTCCGTGCG
 +
|-
 +
| KH20_BBa-delC_rv || 2013-08-09 || submitting delC to registry || GCAT CTGCAGCGGCCGCTACTAGTA TTA TCATGTCGATTCCTTGGTGC
 +
|-
 +
| KH21_BBa-indC_fw || 2013-08-09 || submitting indC to registry || GCAT GAATTCGCGGCCGCTTCTAG ATGTTAGAAAATAATATTACACAATG
 +
|-
 +
| KH22_BBa-indC_rv || 2013-08-09 || submitting indC to registry || GCAT CTGCAGCGGCCGCTACTAGTA TTAGATTATTTTCTCAATCTCAG
 +
|-
 +
| NI01:bpsA_AOxA_PstI_fw|| 2013-06-04 || for amplifying bpsA (AOxA domain) || GAGGAGAAATACTAGATGACACTGCAGGAAACAAGCGTGC
|-
|-
-
| PW13:TycAdE-TycC6_fwd || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI02:bpsA_AOxA_rv|| 2013-06-04 || for amplifying bpsA (AOxA domain)  || GAAGGGCCGTTCCACCAGCTCAGCGTTGACCTGGTCAGAAGCG
|-
|-
-
| IK26:BBa_I746200_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI03:bpsA_T_fw|| 2013-06-04 || for amplifying bpsA (T domain) || GACCAGGTCAACGCTGAGCTGGTGGAACGGCCCTTCGTCG
|-
|-
-
| IK27:BBa_I746200-BBa_B0029_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI04:bpsA_T_rv|| 2013-06-04 || for amplifying bpsA (T domain) || GACGAAGCGACTAGACTCCTGAGCCACTTCTCTCTCCAGCCG
|-
|-
-
| IK28:pccB-BBa_XX_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI05:bpsA_TE_fw|| 2013-06-04 || for amplifying bpsA (TE domain) with RBS 1 || GAGAGAGAAGTGGCTCAGGAGTCTAGTCGCTTCGTCCGA
|-
|-
-
| IK29:accA2-BBa_B0030_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI06:bpsA_TE_Stop_XbaI_rbs_rv|| 2013-06-04 || for amplifying bpsA (TE domain) with RBS 1 || TGGCAGCAGAGCAGCGATCATCTAGTATTTCTCCTCTTTCCTCTAGATCATCATTCCCCCAGCAGGTATCTAAT
|-
|-
-
| IK30:sfp-BBa_B0030-accA2_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI07:svp_fw|| 2013-06-04 || for amplifying svp || GAGGAGAAATACTAGATGATCGCTGCTCTGCTGCCAAGTTGG
|-
|-
-
| IK31:sfp_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI08:svp_rv|| 2013-06-04 || for amplifying svp || GCACTATCAGCGTTATTATGGCACGGCAGTCCTATCGTCG
|-
|-
-
| IK32:BBa_J04450-sfp_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI09:pSB1C3_fw|| 2013-06-04 || for linearizing, amplifying pSB1C3 || GATAGGACTGCCGTGCCATAATAACGCTGATAGTGCTAGTGTAGATCGC
|-
|-
-
| IK33:BBa_XX_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI10:pSB1C3_PstI_rv|| 2013-06-04 || for linearizing, amplifying pSB1C3 || GCTTGTTTCCTGCAGTGTCATCTAGTATTTCTCCTCTTTCTCTAGTATGTG
|-
|-
-
| IK34:pccB_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI11:bpsA_rvN|| 2013-07-15 || for amplifying bpsA (TE domain) with RBS 2 || TGGCAGCAGAGCAGCGATCATTATTTAGGTTTCCTGTGTGAATCATCATTCCCCCAGCAGGTATCTAAT
|-
|-
-
| IK35:BBa_J04450-pccB_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI12:svp_fwN|| 2013-07-15 || for amplifying svp with RBS 2  || CAGGAAACCTAAATAATGATCGCTGCTCTGCTGCCAAGTTGG
|-
|-
-
| RB23:bpsA-fus_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| NI13_ccdb_fw || 2013-07-23 || amplifing ccdb with RBS from pDONR || CGTCGCACCTAGGACCGAAACA GAGAACAGGGGCTGGTGAAATGC
 +
|-
 +
| NI14_ccdb_rv || 2013-07-23 || amplifing ccdb with RBS from pDONR || GCCACTTCTCTCTCCAGCCGTCG TTATATTCCCCAGAACATCAGGTTAATGGCG
 +
|-
 +
| NI15_bpsA_backbone-withoutT_fw || 2013-07-23 || linearization of vector with partial indigoidine synthase || CGACGGCTGGAGAGAGAAGTGGC
 +
|-
 +
| NI16_bpsA_backbone-withoutT_rv || 2013-07-23 || linearization of vector with partial indigoidine synthase || TC TGTTTCGGTCCTAGGTGCGACG
 +
|-
 +
| NI17_indC_T_fw || 2013-07-23 || introducing T domain of indC || CGTCGCACCTAGGACCGAAACA GAAATAAGGCTTGGAAAAATTTGGATGGAAGT
 +
|-
 +
| NI18_indC_T_rv || 2013-07-23 || introducing T domain of indC || GCCACTTCTCTCTCCAGCCGTCG AGCCAATTCTGCTATATTAGGAGATTGA
 +
|-
 +
| NI19_bpsA_T_fw || 2013-07-23 || introducing T domain of bpsA || CGTCGCACCTAGGACCGAAACA GAGAAGGAAATCGCAGCCGTGTGGG
 +
|-
 +
| NI20_bpsA_T_rv || 2013-07-23 || introducing T domain of bpsA || GCCACTTCTCTCTCCAGCCGTCG GGCCAGCTTTTCAATTGTTGGTGACTCC
 +
|-
 +
| NI19:ngrA_fw || || || GAATTCGCGGCCGCTTCTAG ATGGAACAGACAGTTATACACACC
 +
|-
 +
| NI20:ngrA_rv || || || CTGCAGCGGCCGCTACTAGTA TTATTGAATAATTAGCGTTAATATTTCCTG
 +
|-
 +
|RB05:RBS1-BpsA_fw||2013-07-01||Gibson assembly of pSB1C3 bpsA svp||AAAGAGGAGAAA TACTAG ATGACACTTCAGGAAACCAGCG
|-
|-
-
| RB24:bpsA-fus_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB06:BpsA-RBS2_rv||2013-07-01||Gibson assembly of pSB1C3 bpsA svp||CTTTCCTGTGTGA AAGCTT TCA TCACTCCCCGAGCAGATATCG
|-
|-
-
| RB25:svp-fus_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB07:RBS2-svp_fw||2013-07-01||Gibson assembly of pSB1C3 bpsA svp||CTTTCACACAGGAAAG TAAATA ATGGCTGCTCTTCTTCCTAGTTGGGC
|-
|-
-
| RB26:svp-fus_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB08:svp-pSB1C3_rv||2013-07-01||Gibson assembly of pSB1C3 bpsA svp||CTATCAGCGTTATTA AAGCTT TCATCA TGGCACGGCAGTCCTATCG
|-
|-
-
| RB29:svp-Svert_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB09:pSB1C3_fw||2013-07-01||Gibson assembly of pSB1C3 bpsA svp||AAGCTT TAATAACGCTGATAGTGCTAGTGTAGATCGC
|-
|-
-
| RB30:svp-Svert_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB10:pSB1C3-RBS1_rv||2013-07-01||Gibson assembly of pSB1C3 bpsA svp||CTAGTA TTTCTCCTCTTT CTCTAGTATGTGTG
|-
|-
-
| RB31:Sc-indC-Schr_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB11:BpsA_Taka_fw||2013-07-08||colony PCR bpsA Takahashi||cat atgactcttcaggagaccagcgtgctc
|-
|-
-
| RB32:Sc-indC-Schr_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB12:BpsA_Taka_rv||2013-07-08||colony PCR bpsA Takahashi||aag cttctcgccgagcaggtagcggatgtg
|-
|-
-
| RB33:entD-MG1655_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB13:svp_Taka_fw||2013-07-08||colony PCR svp Takahashi||cat atgatcgccgccctcctgccctcctg
|-
|-
-
| RB34:entD-MG1655_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB14:svp_Taka_rv||2013-07-08||colony PCR svp Takahashi||ctcgagcgggacggcggtccggtcgtccgc
|-
|-
-
| RB39:Svert-extr_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB15:svp_Sanch_fw||2013-07-08||PCR svp Sanchez||TATA ATGCATGCTCGCCGCCCTCCCC
|-
|-
-
| RB40:Svert-extr_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB16:svp_Sanch_rv||2013-07-08||PCR svp Sanchez||TTAAGATCTCGGGACGGCGGTCCGGTC
|-
|-
-
| RB41:MG1655-extr_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB17:sfp_fw||2013-07-08||sfp extraction from BAP1||ATGAAGATTTACGGAATTTATATGG
|-
|-
-
| RB42:MG1655-extr_rv || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB18:sfp_rv||2013-07-08||sfp extraction from BAP1||TTATAAAAGCTCTTCGTACGAGACC
|-
|-
-
| RB45:Sc-indC-Schr_rvG || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB19:entD_fw||2013-07-08||entD extraction from E.coli Lambalot||TAAATA ATGGTCGATATGAAAACTACGC
|-
|-
-
| HM_11:lacI_RBS(1)_DelH_rev || Sigma-Aldrich Chemie GmbH || VC00021
+
|RB20:entD_rv||2013-07-08||entD extraction from E.coli Lambalot||AAGCTT ATTAATCGTGTTGGCACAGCG
 +
|-
 +
| RB21:pSB1C3_fw || 2013-07-15 || indigoidine exchangeable construct || TAATGA GCTAGC TAATAACGCTGATAGTGCTAGTG
 +
|-
 +
| RB22:pSB1C3_rv || 2013-07-15 || indigoidine exchangeable construct || CAT GGTACC TTTCTCCTCTTT CTCTAGTATGTGTG
 +
|-
 +
| RB23:bpsA-fus_fw || 2013-07-15 || indigoidine exchangeable construct || CACACATACTAGAG AAAGAGGAGAAA GGTACC ATGACTAGTACACTGCAGG
 +
|-
 +
| RB24:bpsA-fus_rv || 2013-07-15 || indigoidine exchangeable construct || CAT GGATCC GGTTTCCTGTGTGAA TCATTA TTCCCCCAGCAGGTATCTAATATG
 +
|-
 +
| RB25:svp-fus_fw || 2013-07-15 || indigoidine exchangeable construct || CACAGGAAACC GGATCC ATGACTAGTATCGCTGCTCTGCTG
 +
|-
 +
| RB26:svp-fus_rv || 2013-07-15 || indigoidine exchangeable construct || CAGCGTTATTA GCTAGC TCATTA TGGCACGGCAGTCCTATC
 +
|-
 +
| RB27:indC-Plum_fw || 2013-07-15 || indigoidine exchangeable construct || CATACTAGAG AAAGAGGAGAAA GGTACC ATGTTAGAAAATAATATTACACAATG
 +
|-
 +
| RB28:indC-Plum_rv || 2013-07-15 || indigoidine exchangeable construct || CAT GGATCC GGTTTCCTGTGTGAA TTA TTAGATTATTTTCTCAATCTCAG
 +
|-
 +
| RB29:svp-Svert_fw || 2013-07-15 || indigoidine exchangeable construct || CACAGGAAACC GGATCC A TGATCGCCGCCCTC
 +
|-
 +
| RB30:svp-Svert_rv || 2013-07-15 || indigoidine exchangeable construct || CAGCGTTATTA GCTAGC TCA TTACGGGACGGCGGTC
 +
|-
 +
| RB31:Sc-indC-Schr_fw || 2013-07-15 || indigoidine exchangeable construct || GAG AAAGAGGAGAAA GGTACC ATGAGCGTAGAGACCATC
 +
|-
 +
| RB32:Sc-indC-Schr_rv || 2013-07-15 || indigoidine exchangeable construct || CAT AGATCT GGTTTCCTGTGTGAA TTA TCAGTAGTTGGGCGTCTTG
 +
|-
 +
| RB33:entD-MG1655_fw || 2013-07-15 || indigoidine exchangeable construct || CACAGGAAACC GGATCC ATGAAAACTACGCATACCTC
 +
|-
 +
| RB34:entD-MG1655_rv || 2013-07-15 || indigoidine exchangeable construct || CAGCGTTATTA GCTAGC TCA TTAATCGTGTTGGCACAGC
 +
|-
 +
| RB35:sfp-Naka_fw || 2013-07-15 || indigoidine exchangeable construct || CACAGGAAACC GGATCC ATGAAGATTTACGGAATTTATATGGAC
 +
|-
 +
| RB36:sfp-Naka_rv || 2013-07-15 || indigoidine exchangeable construct || CAGCGTTATTA GCTAGC TCA TTATAAAAGCTCTTCGTACGAGAC
 +
|-
 +
| RB37:Plum-extr_fw || 2013-07-15 || genomic extraction || ATGTTAGAAAATAATATTACACAATG
 +
|-
 +
| <div id="RB38">RB38:Plum-extr_rv</div> || 2013-07-15 || genomic extraction || TTAGATTATTTTCTCAATCTCAG
 +
|-
 +
| RB39:Svert-extr_fw || 2013-07-15 || genomic extraction || GTGATCGCCGCCCTC
 +
|-
 +
| RB40:Svert-extr_rv || 2013-07-15 || genomic extraction || TTACGGGACGGCGGTC
 +
|-
 +
| RB41:MG1655-extr_fw || 2013-07-15 || genomic extraction || ATGAAAACTACGCATACCTCCCTC
 +
|-
 +
| RB42:MG1655-extr_rv || 2013-07-15 || genomic extraction || TTAATCGTGTTGGCACAGCGTTATG
 +
|-
 +
| RB43:Bsub-extr_fw || 2013-07-15 || genomic extraction || ATGAAGATTTACGGAATTTATATGGAC
 +
|-
 +
| RB44:Bsub-extr_rv || 2013-07-15 || genomic extraction || TTATAAAAGCTCTTCGTACGAGAC
 +
|-
 +
| RB45:Sc-indC-Schr_rvG || 2013-07-15 || indigoidine exchangeable construct || CAT GGATCC GGTTTCCTGTGTGAA TTA TCAGTAGTTGGGCGTCTTG
|-
|-
-
| HM_12:DelH_RBS(1.2)_mRFP_fw || Sigma-Aldrich Chemie GmbH || VC00021
+
| RB46:indC-woPPT_rv || 2013-08-05 || indC-pSB1C3 w/o PPTase; RB21 overlap || GCACTATCAGCGTTATTA GCTAGCTCATTA TTA TTAGATTATTTTCTCAATCTCAG
 +
|-
 +
| RB47:indC-SpeI_fw || 2013-08-05 || remove cutting sites from indC-Plum || CAAGTTCCTAAACCC ACTAGT CTGGCTTAT ATTATTTATACCTCTGGTAGCAC
 +
|-
 +
| RB48:indC-EcoRI_rv || 2013-08-05 || remove cutting sites from indC-Plum || CAATACCCACC GAATTC TTTGAGCT AATTTCTGACAGACAATACC
 +
|-
 +
| RB49:indC-EcoRI_fw || 2013-08-05 || remove cutting sites from indC-Plum || AGCTCAAA GAATTC GGTGGGTATTG GGCTTTTTTGTGATC
 +
|-
 +
| RB50:indC-SpeI_rv || 2013-08-05 || remove cutting sites from indC-Plum || ATAAGCCAG ACTAGT GGGTTTAGGAACTTG GAACTTGAACTGTG
 +
|-
 +
| RB51:pSB(2/3)K3_fw || 2013-08-05 || PPTase plasmid || TAATGA GCTAGC tactagtagcggccgctgcagtc
 +
|-
 +
| RB52:pSB(2/3)K3_rv || 2013-08-05 || PPTase plasmid || CAT GGATCC GGTTTCCTGTGTGAA ctctagaagcggccgcgaattcc
 +
|-
 +
| RB53:entF-T_fw || 2013-08-07 || bring T-Domain in indC || TGGTGCCATTACATACAGATACT GAAACGATTATCGCCGCGGCATTC
 +
|-
 +
| RB54:entF-T_rv || 2013-08-07 || bring T-Domain in indC || AGAGTCTGTCTGTTCAATCCACTT TGCCAGTTTGGCGACAGTTGACG
 +
|-
 +
| RB55:tycA1-T_fw || 2013-08-07 || bring T-Domain in indC || TGGTGCCATTACATACAGATACT GAATCGATTCTCGTCTCCATCTGG
 +
|-
 +
| RB56:tycA1-T_rv || 2013-08-07 || bring T-Domain in indC || AGAGTCTGTCTGTTCAATCCACTT AGCAACCTGCTCGATCGTCGGGTAATTC
 +
|-
 +
| RB57:tycC6-T_fw || 2013-08-07 || bring T-Domain in indC || TGGTGCCATTACATACAGATACT GAACAGCAACTGGCAGCCATCTGGCAAG
 +
|-
 +
| RB58:tycC6-T_rv || 2013-08-07 || bring T-Domain in indC || AGAGTCTGTCTGTTCAATCCACTT GGCCACCTTTTCGATCGTCGGATACTG
 +
|-
 +
| RB59:delH4-T_fw || 2013-08-07 || bring T-Domain in indC || TGGTGCCATTACATACAGATACT GAGACGCTGCTGGCCCGTATCTG
 +
|-
 +
| RB60:delH4-T_rv || 2013-08-07 || bring T-Domain in indC || AGAGTCTGTCTGTTCAATCCACTT CGCCAGCTCGGCAATGCTTTGC
 +
|-
 +
| RB61:delH5-T_fw || 2013-08-07 || bring T-Domain in indC || TGGTGCCATTACATACAGATACT GCCATGGCGCTGGCCCGCATC
 +
|-
 +
| RB62:delH5-T_rv || 2013-08-07 || bring T-Domain in indC || AGAGTCTGTCTGTTCAATCCACTT CAGCAGGCCGGCAATGGTCG
 +
|-
 +
| RB63:pSB(2/3)K3_rv_korr || 2013-08-07 || PPTase plasmid || CAT GGATCC GGTTTCCTGTGTGAA CTCTAGTATGTGTGAAATTGTTATCC
 +
|-
 +
| RB64:DelC-extr_fw || 2013-08-07 || get Delftia PPTase || ATGCAGCTCGTGTCCGTGCGTGAG
 +
|-
 +
| RB65:DelC-extr_rv || 2013-08-07 || get Delftia PPTase || TCATGTCGATTCCTTGGTGCGCCAC
 +
|-
 +
| RB66:DelC-pSB2K3_fw || 2013-08-07 || DelC in PPTase-construct || CACAGGAAACC GGATCC ATGCAGCTCGTGTCCGTGCGTGAG
 +
|-
 +
| RB67:DelC-pSB2K3_rv || 2013-08-07 || DelC in PPTase-construct || CAGCGTTATTA GCTAGC TCA TCATGTCGATTCCTTGGTGCGCCAC
 +
|-
 +
| RB68:indC-SpeI_fw_corr || 2013-08-09 || indC w/o cutting sites corrected || CAAGTTCCTAAACCC ACgAGT CTGGCTTAT ATTATTTATACCTCTGGTAGCAC
 +
|-
 +
| RB69:indC-EcoRI_rv_corr || 2013-08-09 || indC w/o cutting sites corrected || CAATACCCACC GAAcTC TTTGAGCT AATTTCTGACAGACAATACC
 +
|-
 +
| RB70:indC-EcoRI_fw_corr || 2013-08-09 || indC w/o cutting sites corrected || AGCTCAAA GAgTTC GGTGGGTATTG GGCTTTTTTGTGATC
 +
|-
 +
| RB71:indC-SpeI_rv_corr || 2013-08-09 || indC w/o cutting sites corrected || ATAAGCCAG ACTcGT GGGTTTAGGAACTTG GAACTTGAACTGTG
 +
|-
 +
| RB72:bpsA-TTE_rv || 2013-08-09 || exchange T and TE-domain || GCACTATCAGCGTTATTA GCTAGC TCATTA TTCCCCCAGCAGGTATCTAATATG
 +
|-
 +
| RB73:entF-TTE_rv || 2013-08-09 || exchange T and TE-domain || GCACTATCAGCGTTATTA GCTAGC TCA TTACCTGTTTAGCGTTGCGCGAATAATCG
 +
|-
 +
| RB74:tycC6-TTE_rv || 2013-08-09 || exchange T and TE-domain || GCACTATCAGCGTTATTA GCTAGC TCA TTATTTCAGGATGAACAGTTCTTGC
 +
|-
 +
| RB75:delH5-TTE_rv || 2013-08-09 || exchange T and TE-domain || GCACTATCAGCGTTATTA GCTAGC TCATTA TCAGTCCAGCGCGTACTCCAG
|-
|-
 +
| RB76:plu2670-T_fw || 2013-08-15 || bring T-Domain in indC || TGGTGCCATTACATACAGATACT GAAACCACACTGGCTGCTATC
 +
|-
 +
| RB77:plu2670-T_rv || 2013-08-15 || bring T-Domain in indC || AGAGTCTGTCTGTTCAATCCACTT CGCAAATGCGGATAACACC
 +
|-
 +
| RB78:indC-SpeI_rv_corr2 || 2013-08-26 || indC w/o cutting sites w/o frameshift || ATAAGCCAG ACTcGT GGGTTTAGGAACTTG AACTGTG
 +
|-
 +
| RB79:indC_woPPTase_rv2 || 2013-08-29 || indC-pSB1C3 w/o PPTase; RB35 5' part || CAGCGTTATTA GCTAGC TCA TTAGATTATTTTCTCAATCTCAG
 +
|-
 +
| RB80:indCdT_3_fw || 2013-09-06 || T-domain border combinations bpsA || GGCGGATATCCTATGAGTTTGAGATTGC
 +
|-
 +
| RB81:indCdT_3_rv || 2013-09-06 || T-domain border combinations bpsA || TCCATTGGCCGTCAAAGGTAATTTATCG
 +
|-
 +
| RB82:bpsA-T3_fw || 2013-09-06 || T-domain border combinations bpsA || CGATAAATTACCTTTGACGGCCAATGGA AAGATCGATGTGAAAGCACTGGCCGCTTCTGACC
 +
|-
 +
| RB83:bpsA-T3_rv || 2013-09-06 || T-domain border combinations bpsA || GCAATCTCAAACTCATAGGATATCCGCC CAGACCTGGCCAGCAGATCACAGG
 +
|-
 +
| RB84:indCdT_4_fw || 2013-09-06 || T-domain border combinations bpsA || TTTGAAGTTGCATACCAGCTTGAACAAGC
 +
|-
 +
| RB85:bpsA-T4_rv || 2013-09-06 || T-domain border combinations bpsA || GCTTGTTCAAGCTGGTATGCAACTTCAAA TGCCACGCGAGCTCCGAAGCTATATCC
 +
|-
 +
| RB86:indC-T2_fw || 2013-09-06 || T-domain border combinations bpsA || TTACAGATACTTTTTCAATCTCCTAATATAGCAG
 +
|-
 +
| RB87:indC-T2_rv || 2013-09-06 || T-domain border combinations bpsA || TACTTCCATCCAAATTTTTCCAAGC
 +
|-
 +
| RB88:bpsA-T2_fw || 2013-09-06 || T-domain border combinations bpsA || GCTTGGAAAAATTTGGATGGAAGTA CTGAGACGCGAAAATGCTAGTGTCC
 +
|-
 +
| RB89:bpsA-T2_rv || 2013-09-06 || T-domain border combinations bpsA || TTAGGAGATTGAAAAAGTATCTGTAA AGGCAGGGACACTCCCAGTCTAGCATTCAG
 +
|-
 +
| RB90:indC-T-screen_fw || 2013-09-13 || screeening primer || GATCAATGCGGCCTTTAATATTCG
 +
|-
 +
| RB91:bpsA-T-screen_fw || 2013-09-13 || screeening primer || GGAACTGAATGCTAGACTGGGAG
 +
|-
 +
| RB92:entF-T3_fw || 2013-09-16 || native T with border combination 31 || CGATAAATTACCTTTGACGGCCAATGGA AAGCTGGATCGCAAAGCCTTACCGTTG
 +
|-
 +
| RB93:tycA1-T3_fw || 2013-09-16 || native T with border combination 31 || CGATAAATTACCTTTGACGGCCAATGGA AAGATCGACCGCAAAGCGTTGC
 +
|-
 +
| RB94:tycC6-T3_fw || 2013-09-16 || native T with border combination 31 || CGATAAATTACCTTTGACGGCCAATGGA AAAGTGGATCGCAAAGCTTTG
 +
|-
 +
| RB95:delH4-T3_fw || 2013-09-16 || native T with border combination 31 || CGATAAATTACCTTTGACGGCCAATGGA AAGCTGGACCGGCAGGCCCTG
 +
|-
 +
| RB96:delH5-T3_fw || 2013-09-16 || native T with border combination 31 || CGATAAATTACCTTTGACGGCCAATGGA AAGCTTGACCGTGGTGCGCTG
 +
|-
 +
| RB97:plu2642-T3_fw || 2013-09-16 || native T with border combination 31 || CGATAAATTACCTTTGACGGCCAATGGA AAAATCGATTTCGACACATTACAAG
 +
|-
 +
| RB98:plu2670-T3_fw || 2013-09-16 || native T with border combination 31 || CGATAAATTACCTTTGACGGCCAATGGA AAGCTGGACCGTCGGGCGTTACCGGCAC
 +
|-
 +
| RB99:plu2642_pRB23-plu_fw || 2013-09-24 || plu2642 on pSB1C3 || CATACTAGAG AAAGAGGAGAAA GGTACC ATGCAATCAACTCTCCCAATAATAAAATGG
 +
|-
 +
| RB100:plu2642_pRB23-plu_rv || 2013-09-24 || plu2642 on pSB1C3 || CAGCGTTATTA GCTAGC TCA TCAACTCAAGAATCGAGCTAATTCGTTAACAC
 +
|-
 +
| RB101:plu2642_A_rv || 2013-09-24 || shuffling of all domains || GTTCTGGCATCCTTCTTAATTAACAT TGCAGCGTGTTTCACTTGATCGATAAC
 +
|-
 +
| RB102:indC_Ox_fw || 2013-09-24 || shuffling of all domains || ATGTTAATTAAGAAGGATGCCAGAAC
 +
|-
 +
| RB103:plu2642_valInd_fw || 2013-09-24 || shuffling of all domains || CAGCCTGCCAGCAGCAAAACG ATGCAATCAACTCTCCCAATAATAAAATGG
 +
|-
 +
| RB104:plu2642_valInd_rv || 2013-09-24 || shuffling of all domains || CAT CGTTTTGCTGCTGGCAGGCTG
 +
|-
 +
| RB105:tycC2_T3_fw || 2013-09-24 || shuffling of all domains || CGATAAATTACCTTTGACGGCCAATGGA AAAGTGGATCGCAAGGCATTG
 +
|-
 +
| RB106:tycC2_T1_rv || 2013-09-24 || shuffling of all domains || AGAGTCTGTCTGTTCAATCCACTT GGCCAGGCCCGCGATCGTTGG
 +
|-
 +
| RB107:tycC2_A_fw || 2013-09-24 || shuffling of all domains || CATACTAGAG AAAGAGGAGAAA GGTACC ATGACGGAAGCGGAAAAACGCACACTCCT
 +
|-
 +
| RB108:tycC2_A_rv || 2013-09-24 || shuffling of all domains || GTTCTGGCATCCTTCTTAATTAACAT GACGACCGCTTTGACCGCTTCATG
 +
|-
 +
| RB109:tycC2_valInd_fw || 2013-09-24 || shuffling of all domains || CAGCCTGCCAGCAGCAAAACG ATGACGGAAGCGGAAAAACGCACACTCC
 +
|-
 +
| RB110:plu2642_TE_fw || || shuffling of all domains || AGTGGATTGAACAGACAGACTCT AAAATCGTTGAAGGTGAAGTAACCAG
 +
|-
|}
|}

Revision as of 21:57, 4 October 2013

Contents

Materials used during the project

Partly adopted from iGEM Team Heidelberg 2009 and 2010. Please recheck.

Table 1: Kits

Kit Supplier Catalog Number
QIAquick® Gel Extraction Kit (250) QIAGEN 28706
QIAquick® Nucleotid Removal Kit (250) QIAGEN 28306
QIAEX II® Gel Extraction Kit (500) QIAGEN 20051
MinElute® PCR Purification Kit (250) QIAGEN 28006
QIAquick® PCR Purification Kit (250) QIAGEN 28106
QIAGEN® Plasmid Plus Midi Kit (100) QIAGEN 12945
QIAprep® Spin Miniprep Kit (250) QIAGEN 27106
Plasmid Plus Maxi Kit (25) QIAGEN 12963
Plasmid Plus Midi Kit (25) QIAGEN 12943
QIAprep Spin Miniprep Columns QIAGEN 27115

Table 2: Marker

Marker Supplier Catalog Number
Quick-Load 2-Log DNA Ladder (0.1-10.0 kb) New England BioLabs N3200S
Quick-Load® 1 kb DNA Ladder New England BioLabs N0468S
50 bp DNA Ladder New England BioLabs N3236S
Gel loading solution Sigma-Aldrich Chemie GmbH G2526-5ML

Table 3: Enzymes

Enzyme Supplier Catalog Number
EcoRI New England BioLabs
NotI-HF New England BioLabs R3189 S
PstI New England BioLabs
PstI-HF New England Biolabs R3140 S
SpeI-HF New England BioLabs R3133 S
XbaI New England BioLabs R0145 S
PvuI-HF New England BioLabs R3150S
NheI-HF New England BioLabs R3131 S
KpnO-HF New England BioLabs
Phusion Flash High-Fidelity PCR MAster Mix Biozym Scientific GmbH F-548L
Phusion® High-Fidelity PCR Master mix New England Biolabs GmbH M0531 L
Gibson Assembly# Master Mix New England Biolabs GmbH E2611 S
Lambda DNA Mono Cut Mix New England Biolabs GmbH N3019 S
T4 DNA Ligase New England Biolabs GmbH M0202 S
PacI, recombinant New England Biolabs GmbH R0547 S
EcoRI-HF New England Biolabs GmbH R3101 S
SalI-HF New England Biolabs GmbH R3138 S
NotI-HF New England Biolabs GmbH R3189 S
SpeI-HF New England Biolabs GmbH R3133 S
2x PCR Master mix Solution (iTaq) HISS DIAGNOSTICS GmbH 25028
DpnI New England Biolabs GmbH R0176 S
KpnI-HF New England Biolabs GmbH R3142 S
Calcium Carbonate Bioxtra Sigma-Aldrich Chemie GmbH C4830-100G
BamI New England Biolabs GmbH R3136 S
HindIII-HF New England Biolabs GmbH R3104 S
Dream Taq PCR MM Fermentas Life Sciences K1071
Lysozyme from chicken egg white Sigma-Aldrich Chemie GmbH L4919-500MG
Pronasw E from Streptomyces griseus Sigma-Aldrich Chemie GmbH P6911-100MG
BgIII New England Biolabs GmbH R0144 S
Gibson Assembly# Master Mix New England Biolabs GmbH E2611 S
MfeI-HF New England Biolabs GmbH R3589 S
DreamTaq Green PCR Master Mix (2X) Thermo FisherScientificBiosciences GmbH K1081

Table 4: Bacteria

Strain Supplier Catalog Number
DH10beta
Top10
BapI
Delftia acidovorans DSMZ DSM39
Streptomyces lavendulae lavendulae DSMZ DSM40708
Streptomyces mobaraensis DSMZ DSM 40903
Photorhabdus laumondii luminescens DSMZ DSM 15139
Delftia acidovorans DSMZ DSM 14801

Table 5: Media and Antibiotics

Antibiotic Supplier Catalog Number Final Concentration↓ Solvent
Ampicillin anhydrous crystalline Sigma-Aldrich Chemie GmbH A9393-5G 1:1000 Ethanol
Ampicillin sodium crystalline A9518-5G 1:1000 Ethanol
Chloramphenicol crystalline Sigma-Aldrich Chemie GmbH C0378-5G 1:3000 Ethanol
Kanamycinsulfat Gemisch der Komponentena Sigma-Aldrich Chemie GmbH 60615-5G 1:1000 H2O
Tetracycline
SOC Outgrowth Medium New England Biolabs GmbH B9020 S
LB BROTH BASE Th. Geyer GmbH & Co KG SA/L3022/001000 20g/1l H2O
LB Broth powder Sigma-Aldrich Chemie GmbH L3022-1KG
Agarose molecular Biology Reagent Th. Geyer GmbH & Co KG SA/A9539/000050 0,5% H2O
Propionic acid sodium insect cell*culture Sigma-Aldrich Chemie GmbH P5436-100G
Agarose for routine use Sigma-Aldrich Chemie GmbH A9539-100G
Bacitracin Sigma-Aldrich Chemie GmbH B0125-50KU

preparation of M9 minimal media according to Serva company

Table 6: Primers and Oligos

Misc Primers

Identifier Order date Note Sequence
VF2 - for screening in standard BB backbone- binds on the Backbone before Insert tgccacctgacgtctaagaa
VR - for screening in standard BB backbone- binds on Backbone behind Insert attaccgcctttgagtgagc

Delftibactin

Identifier Order date Note Sequence
DN01:delH_f1_PacI_fw - fw Primer for DelH-fragment1 with RBS and PacI-restriction site tttt ttaattaa tcacacaggaaagtactag ATGGACCGTGGCCGCCTGC GCCAAATCG
DN02:delH_f1_SalI_rev - - tttt GTCGACCAACACCTGTGCCTGC
DN03:delH_f2_SalI_fw - - tttt GTCGACTGGATGGAGCCTGGTGAAAG
DN04:delH_f2_KpnI_rev - - tttt ggtacc TCAGTCCAGCGCGTACTCCAG
DN05:AraCbb_KpnI_fw - amplifying the Backbone for DelH (pSB6A1-AraC-lacZ) tttt ggtacc aaagaggagaaatactagatgaccatg
DN06:AraCbb_PacI_rev2 15-05-2013 amplifying the Backbone for DelH (pSB6A1-AraC-lacZ) tttt ttaattaa gctagcccaaaaaaacggtatg
DN07:Screen_delH_rev 15-05-2013 for screening if DelH is present - binds on the very beginning of DelH CTTTCCTCGAACACCGTGCGCAG
DN08:DelH_EcoRI_rev - rev_Primer for DelH Fragment f1a CTCGTCGCCATGGACCAGGCAG
DN09:DelH_f1_fw_long 2013-06-11 for amplifying DelH-1a from the genome: doesn't work ATGGACCGTGGCCGCCTGCGCCAAATCG
DN10:DelH_f1_fw_short 2013-06-11 for amplifying DelH-1a from the genome: doesn't work ATGGACCGTGGCCGCCTGC
DN11:DelH_f1_fw_short2 2013-06-11 for amplifying DelH-1a from the genome: works!!!! GCCGCCTGCGCCAAATCG
DN12:DelH_f1_PacI_fw_short 2013-06-11 for amplifying DelH-1a from the genome: doesn't work ttttttaattaatcacacaggaaagtactagATGGACCGTGGCCGCCTGC
DN13:Screen_DelH_fw 15-05-2013 PCR Screening for presence of DelH insert gtaaacccactggtgataccattc
FS_01: pSB4K5_DelA_rv20-13-06-28Amplification of pSB4K5 from the iGEM Distribution Gibson Primer with overhang to DelA introducing the RBS BBa_B0035 TCGCGGCGATCCGGTACTGCGCCTCTGTTGAACATCTGATATTCTCCTCTTTAATCGACAGATTGTGTGAAATTGTTATCCGCTCAC
FS_02: DelAG_1_fw2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer TTCAACAGAGGCGCAGTACCGGATC
FS_03: DelAG_1_rv2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer GTCGGAGACGATGTGGTGCATCAC
FS_04: DelAG_2_fw2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer CTGCAGGCCAATGAGCACATCCTG
FS_05: DelAG_2_rv2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer CACAGGTGGTAGATGGCGTC
FS_06: DelAG_3_fwG2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer ATTGCGAGGACTTGCTCGATG
FS_07: DelAG_3_rv2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer TTTGCTGCAGCGCCAGCACATCGAG
FS_08: DelAG_4_fw2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer GTACGGCCTATCACATCAGCG
FS_09: DelAG_4_rv2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer GAAGCTCAGCAGGTTGGGCGAGACG
FS_10: DelAG_5_fw2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer GAATTTTGTTCCACCACCTGCTG
FS_11: DelAG_5_rv2013-06-28 Amplification of DelAG from Delftia acidovorans genome Gibson Primer with overhang to DelOP CTTGAGCAGGCGCAGTACCTCGGAGGGCGGTCGGCTGGCGTTTTCCATGATTCAGGTTTCCTGTGTGAAGCTCATCTCAGATATCTCCCAGAGTTTCGAGAAAG
FS_11: DelAG_5_short_rv2013-05-07 Amplification of DelAG from Delftia acidovorans genome Gibson Primer TCAGATATCTCCCAGAGTTTCGAGAAAG
FS_12: DelOP_fw2013-06-28 Amplification of DelOP from Delftia acidovorans genome Gibson Primer GAATCATGGAAAACGCCAGCCGAC
FS_13: DelOP_rv2013-06-28 Amplification of DelOP from Delftia acidovorans genome Gibson Primer with overhang to DelL CAATGTTGGAGGGGCCGAAGCCGATGCCGATCAGCGGGTGGGTTTGCATGGAAGGTCCTTTCATTGGGTCGATGCGTCCAGTGTCACACCGTGGTGTCTGCAGGCG
FS_13: DelOP_short_rv2013-05-07 Amplification of DelOP from Delftia acidovorans genome Gibson Primer with overhang to DelLTCACACCGTGGTGTCTGCAGGCG
FS_14: DelL_fw2013-06-28 Amplification of DelL from Delftia acidovorans genome Gibson Primer CAAACCCACCCGCTGATCGGCATC
FS_15: DelL_mRFP_pSB4K5_rv2013-06-28 Amplification of DelL Gibson Primer with overhang to BBa_J04450 GAAACGCATGAACTCTTTGATAACGTCTTCGGAGGAAGCCATCTAGTATTTCTCCTCTTTCTCTAGTATCAGTCCTGCAGCGCCAGCTGTTCTGTG
FS_15: DelL_mRFP_pSB4K5__short_rv2013-05-07 Amplification of DelL Gibson Primer with overhang to BBa_J04450TCAGTCCTGCAGCGCCAGCTGTTCTGTG
FS_16: mRFP_pSB4K5_fw (Del)2013-06-28 Amplification of pSB4K5 from iGEM Distribution Gibson Primer GCTTCCTCCGAAGACGTTATC
FS_20: DelF_fw2013-07-13 Amplification of DelF from Delftia acidovorans genome Gibson Primer GACTTGCTCGATGCGGTGCAG
FS_21: DelF_fw2013-07-13 Amplification of DelF from Delftia acidovorans genome Gibson Primer GACGCCATCTACCACCTGTG
FS_22: DelOP_short_fw2013-08-02 Amplification of DelOP from Delftia acidovorans genome inlcuding the recently predicted endogenous Promotor for DelOP Gibson Primer GATGACGCAGGGCGGCGGAATTTGTTCATC
FS_23: DelG_long_rv2013-07-13 Amplification of DelG from Delftia acidovorans genome Gibson primer with overhang to DelOP element including the recently predicted endogenous Promotor GATGAACAAATTCCGCCGCCCTGCGTCATCTCAGATATCTCCCAGAGTTTCGAGAAAG
FS_24: DelAE_rv2013-07-13 Amplification of DelAE from Delftia acidovorans genome Gibson Primer CAGAAGAATTCCCAGAAGGAGATGTCGAAG
FS_25: DelEF_fw2013-07-13 Amplification of DelEF from Delftia acidovorans genome Gibson Primer ACACGGTGCTGCAGAAAACGCCCTTC
FS_26: DelFG_rv2013-07-13 Amplification of DelFG from Delftia acidovorans genome Gibson Primer GAATTCATCCACGATGATCTGCATG
FS_27: DelOP_rv2013-08-02 Amplification of DelOP from Delftia acidovorans genome Gibson Primer CTTTGGGCCGTGCCGGTTTTTGAGATAC
FS_28: DelOP_rv2013-08-02 Amplification of DelOP from Delftia acidovorans genome Gibson Primer GTTTTTGAGATACGCGCGTTGTCAC
FS_29: DelOP_rv2013-08-02 Amplification of DelOP from Delftia acidovorans genome Gibson Primer TTCCCCCTCTCTTTCTCGCTTC
FS_30: DelOP_rv2013-08-02 Amplification of DelOP from Delftia acidovorans genome Gibson Primer CCGCTTCCCCCTCTCTTTCTCGCTTC
FS_31: DelOP_fw2013-08-02 Amplification of DelOP from Delftia acidovorans genome Gibson Primer GTTGGCGAGTTCAAGAAATG
FS_32: DelOP_fw2013-08-02 Amplification of DelOP from Delftia acidovorans genome Gibson Primer TCCTTCAGGTGTGCGGCAGACAAG
FS_33: DelOP_fw2013-08-02 Amplification of DelOP from Delftia acidovorans genome Gibson Primer TTCTTCGTGATGACGCAGGGCGGCGGAATTTGTTC
FS_35: DelG_fw2013-08-05 Amplification of DelG from Delftia acidovorans genome Gibson Primer CATGCAGATCATCGTGGATGAATTC
FS_45: pSB4K5_fw2013-08-02 Amplification of pSB4K5 from iGEM Distribution Gibson Primer without mRFP CCAGGCATCAAATAAAACGAAAG
FS_46: DelL_rv 2013-08-02 Amplification of DelL from Delftia acidovorans genome Gibson Primer creating overlap to pSB4K5 without mRFP TCAGTCCTGCAGCGCCAGCTGTTCTGTGCTTTCGTTTTATTTGATGCCT
FS_47_screening_BB_AF_fw2013-08-02 Primer for screening/sequencing of pFSN construct GTTGGCCGATTCATTAATGC
FS_48_screening_BB_AF_rv2013-08-02 Primer for screening/sequencing of pFSN construct TAACGGTATCGGTATCGCTTTG
FS_49_screening_AFI_AFII_fw2013-08-02 Primer for screening/sequencing of pFSN construct GTTTCTCTGGAAGATGGATAC
FS_50_screening_AFI_AFII_rv2013-08-02 Primer for screening/sequencing of pFSN construct GTTGACGAAAAAGCCGACCAC
FS_51_screening_AF_FG(21-26)_fw2013-08-02 Primer for screening/sequencing of pFSN construct TGGATATCGACTGGACTGCCTG
FS_52_screening_AF_FG(21-26)_rv2013-08-02 Primer for screening/sequencing of pFSN construct TGCACCACATCGACGAAACGG
FS_53_screening_FG(21-26)_G_fw2013-08-02 Primer for screening/sequencing of pFSN construct GTACGGCCTATCACATCAGCG
FS_54_screening_FG(21-26)_G_rv2013-08-02 Primer for screening/sequencing of pFSN construct GAACCTGGGTGTTCACGAAAAAGCC
FS_55_screening_G_OP_fw2013-08-02 Primer for screening/sequencing of pFSN construct GTATCTCTACATGCATCGCTAC
FS_56_screening_G_OP_fw2013-08-02 Primer for screening/sequencing of pFSN construct AGGACATTTTCCGCACCCCG
FS_57_screening_G_OP_rv2013-08-02 Primer for screening/sequencing of pFSN construct GCTGGCGTTTTCCATAAG
FS_58_screening_OP_L_fw2013-08-02 Primer for screening/sequencing of pFSN construct GAACAACTTCCAGCACAGCCTGTTC
FS_59_screening_OP_L_rv2013-08-02 Primer for screening/sequencing of pFSN construct CGTTGAAGATTTCGTTGACG
FS_60_screening_L_BB_fw2013-08-02 Primer for screening/sequencing of pFSN construct CATCTTCAAGGTGTTCTATGAAC
FS_61_screening_L_BB(with_mRFP)_rv2013-08-02 Primer for screening/sequencing of pFSN construct CAGTTTAACTTTGTAGATGAAC
FS_66: DelH_rv2013-08-26 Amplification of DelH from Delftia acidovorans genome Gibson Primer TGGGCATTCACCGCATCGATC
FS_67: DelH_fw2013-08-26 Amplification of DelH from Delftia acidovorans genome Gibson Primer CTTCACGTTGATTGCGCATG
FS_68: DelH_rv2013-08-26 Amplification of DelH from Delftia acidovorans genome Gibson Primer CAGAAGAACTCCCAGACCGAC
FS_69: DelH_fw2013-08-26 Amplification of DelH from Delftia acidovorans genome Gibson Primer GACACCGTTCAGCTTCGATG
FS_70: DelH_rv2013-08-26 Amplification of DelH from Delftia acidovorans genome Gibson Primer GAAGCTGCTCCGCTGATAGAT
FS_71: DelH_fw2013-08-26 Amplification of DelH from Delftia acidovorans genome Gibson Primer ATGTGCTGTCGCTCAAGATG
FS_72_SR_02_fw2013-08-30 Screening of pFHFSN ATGTGCTGTCGCTCAAGATG
FS_73_SR_03_fw2013-08-30 Screening of pFHFSN GTGCTGTTTGGCCGTATG
FS_74_SR_04_fw2013-08-30 Screening of pFHFSN ATCAGGTGCTGAGCTACGAC
FS_75_SR_05_fw2013-08-30 Screening of pFHFSN CTGTTCATCAACACCTTGCC
FS_76_SR_06_rv2013-08-30 Screening of pFHFSN GAAGACAGTCATAAGTGCGGC
FS_77_rv 2013-09-11 Gibson-Primer rev, Amplficiation of the Backbone pSB6A1 with overlap to the RBS BBa_B0034 and the lacI-promotor, it creates an overlap to the beginning of DelH GCGATTTGGCGCAGGCGGCCACGGTCCATCTAGTATTTCTCCTCTTTC
FS_78_rv 2013-09-26 Gibson-Primer rev, Amplficiation of the Backbone pSB6A1 introducing the RBS BBa_B0032 and the promotor BBa_J23114 and creating an overlap to the first fragment of DelH amplified with primer DN_11 GATTTGGCGCAGGCGGCCACGGTCCATCTAGTACTTTCCTGTGTGACTCTAGAGCTAGCATTGTACCTAGGACTGAGCTAGCCATAAACTCTAGAAGCGGCCGCGAATTC
FS_84_fw 2013-09-26 Gibson-Primer fw, Amplficiation of the first fragment of DelH introducing the RBS BBa_B0032 and creating an overlap to primer FS_85 thereby partially introducing the promotor BBa_J23114 GCTCAGTCCTAGGTACAATGCTAGCTCTAGAGTCACACAGGAAAGTACTAGATGGACCGTGGCCGCCTGCG
FS_85_rv 2013-09-26 Gibson-Primer rev, Amplficiation of the Backbone pSB6A1, partially introducing the promotor BBa_B0032 with overlap to primer FS_84 and therefore the promotor BBa_J23114, it creates an overlap to the beginning of DelH GCGATTTGGCGCAGGCGGCCACGGTCCATCTAGTATTTCTCCTCTTTC
FS_86_rv 2013-09-27 Gibson-Primer rev, Amplficiation of the Backbone pSB4K5 without any promotor introducing a KpnI cutting site for restriction cloning, creates an overlap to DelH and will be used for the ccdB strategy GGCGATTTGGCGCAGGCGGCCACGGTCCATGTACTTCGAGTCACTAAGGGCTAAC
FS_87_fw 2013-09-27 Gibson-Primer fw, Amplficiation of the Backbone pSB6A1 introducing a BamHI cutting site for restriction cloning and creating an overlap to the last fragment of DelH CGCTGGAGTACGCGCTGGACTGAGATCCCAGGCATCAAATAAAACG
FS_90_fw 2013-09-27 Gibson-Primer fw, Amplficiation of the ccdB cassette from the template pDonorPlasmid introducing a KpnI cutting site for restriction cloning, creates an overlap to the promotor BBa_J23114 and will be used for the ccdB strategy CTCAGTCCTAGGTACAATGCTAGCTCTAGAGTCACACAGGAAAGCAGTACACTGGCTGTGTATAAGGGAG
FS_93_rv 2013-09-27 Gibson-Primer rev, Amplficiation of the ccdB cassette from the template pDonorPlasmid introducing a BamHI cutting site for restriction cloning, creates an overlap to the backbone pSB6A1 and will be used for the ccdB strategy GTTCACCGACAAACAACAGATGATCCGCGTGGATCCGGCTTAC
FS_94_fw 2013-09-27 Primer fw, Amplficiation of the backbone pSB6A1, will be used for the ccdB strategy ATCTGTTGTTTGTCGGTGAACGC
HM01:DelH_EcoRI_fw - fw_Primer for DelH Fragment f1b GCATTGGAGCCTCAATGGCAAGTC
HM02:DelH_Gib1.1_rev2013-07-09 Gibson-Primer DelH TGCTGCGCCTGCATACGGCCAAACA
HM03:DelH_Gib1.2_fw2013-07-09 Gibson-Primer DelH AGCGGCAGGGACGACGTGGT
HM04:DelH_Gib1.2_rev2013-07-09 Gibson-Primer DelH CATAGAGGTTGTAGAGA
HM05:DelH_Gib2.1_fw2013-07-09 Gibson-Primer DelH AGAACGCCGTCTTCAGGCTCCTG
HM06:DelH_Gib2.1_rev2013-07-09 Gibson-Primer DelH CAATGCTTTG CCGCTCGAA
HM07:DelH_Gib2.2_fw2013-07-09 Gibson-Primer DelH TCGCCACGGCAGCTGTTCGA
HM08:DelH_Gib2_end_rev2013-07-09 Gibson-Primer DelH TCAGTCCAGCGCGTACTCCAG
HM09:AraC_RBS_Delh_rev2013-07-09Gibson-Primer rev, introduces a new RBS and has the AraC-promotor and the beginning of DelH TTGCAAAGCGCTCGGCGATTTGGCGCAGGCGGCCACGGTCCATttaactTTCTCCTCTTTAATactttgagctagcccaa

Aaaaacggtatggagaaacagtagagagtt

HM10:RBS_lacZ2013-07-09 Gibson-Primer fw for the pSB6A1 Backbone with the end of DelH, RBS(1) and the beginning of lacZ TGGAGTACGCGCTGGACTGA TCTAGAG AAAGAGGAGAAA TACTAG ATGACCATGATTA
HM11:lacI_RBS(1)_DelH_rev 2013-07-24 Gibson-Primer rev, amplify the Backbone with overlap with the RBS and the lacI-promotor and it creates and overlap to the start of DelH TCGGCGATTTGGCGCAGGCGGCCACGGTCCATctagtatttctcctctttctctagtatgtgtg
HM12:DelH_RBS(1.2)_mRFP_fw 2013-07-24 Gibson-Primer fw for the pSB6A1 Backbone with the end of DelH, introducing a new RBS(new) and the beginning of mRFP ATTGGCGCTGGAGTACGCGCTGGACTGAtcaaagtATTAAAGAGGAGAAagttaaatggcttcctccgaagacgttatcaAagag
HM13:Screen_DelH_end_fw2013-08-16 New screening primer for the end of DelH together with the VR2 primer from the registry TTTCTGACGACCCTGCACCTGAAG
HM14:DelH_tetR_fw2013-08-16 Gibson-Primer DelH-tetR: amplifies the tetracycline resistance from the pSB1T3 Backbone and creates an overlap to the end of DelH ATTGGCGCTGGAGTACGCGCTGGACTGA atgaagttttaaatcaatctaaag
HM15:tetR_stop_BB_rev2013-08-16 Gibson-Primer tetR-pSB6A1: amplifies the tetracycline resistance and creates an overlap with the Terminator of the Backbone pSB6A1 Cgactgagcctttcgttttatttgatgcctggc ctcgtgatacgcctatttttatagg
HM16:tetR_pSB6A1_fw2013-08-16Gibson-Primer DelH, amplifies the Backbone pSB6A1 creating an overlap with the tetracycline resistance Aaaaataggcgtatcacgag gccaggcatcaaataaaacgaaaggctcag
HM17:DelH_Terminator_BB_fw 2013-08-16 Gibson-Primer fw for the pSB6A1 Backbone (binding the terminator) and creating an overlap with the end of DelH ATTGGCGCTGGAGTACGCGCTGGACTGA aggcatcaaataaaacgaaaggctcag
HM20:BB_HPLC_rev 11-09-2013 HPLC version of HM11
Gibson-Primer rev, amplify the Backbone with overlap with the
RBS and the lacI-promotor and it creates and overlap to
the start of DelH 		GATTTGGCGCAGGCGGCCACGGTCCATctagtatttctcctctttc
HM21:fw_lacI_BbsI_Xba 2013-09-15 Forward primer for cutting out mutated fragment for mutagenesis TTTTGAAGACAA CTAGGCAATACGCAA
HM22:rev_RBS 2013-09-15 Reverse Primer in RBS for mutagenesis TTTTGAAGACAA CTCTTTCTCTAGTATGTGTGAAATTG
HM23:fw_RBS 2013-09-15 Forward Primer in RBS for mutagenesis TTTTGAAGACAA AGAGGAGAAATACTAGATGGACCGTGGC
HM24:rev_BbsI_MfeI 2013-09-15 Reverse primer for cutting out mutated fragment for mutagenesis TTTTGAAGACAA AATTGGACAGCGCGGCATGCCGGTTG
IK01:pLF03_integr_argK_fw 2013-06-12 For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome. GCTGATGGAAGTGGCTGATCTGATC
IK02:pLF03_integr_pET21c_rev 2013-06-12 For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against pLF03 backbone (pET-21c). TCCGCTCACAATTCCCCTATAGTG
IK03:pLF03_integr_argK_rev 2013-06-18 For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome for positive control (to be used with IK01 or IK04). GATAAATTCACTGAGCTGCCGCAG
IK04:pLF03_integr_argK_fw 2013-06-18 For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome. Alternative forward primer in case IK01 does not work. GCGGGAATTAATGCTGTTATGCGAAG
IK05:pLF03_integr_ygfG_fw 2013-06-18 For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome, namely ygfG, which should be replaced by the construct. GCGGTCATTGAGTTTAACTATGGCC
IK06:pLF03_integr_ygfG_rev 2013-06-18 For verification of correct genomic integration of pLF03 into BAP1 via colony-PCR. Primer against BAP1 genome, namely ygfG, which should be replaced by the construct. AACATGGTTGAGGATGCCGACAGC
IK07:ygfG21C1 2013-06-25 For insertion of methylmalonyl-CoA synthesis pathway into E. coli. Extended ygfG21C1 with higher melting temperature. TCACCGCGCACCGGCCTGCGGCAGCTCAGTGAATTTATCC AGATCTCGATCCCGCGAAATTAATAC
IK08:ygfG21C2 2013-06-25 For insertion of methylmalonyl-CoA synthesis pathway into E. coli. Extended ygfG21C2 with higher melting temperature. GTTATGCTGGATAATTTCTGCCGCTTCATTGGCGGTCATC CAAAAAACCCCTCAAGACCCGTTTAG
IK24:pccB_fw 2013-07-16 Colony-PCR of methylmalonyl-CoA synthesis pathway CGTATCGAGGAAGCGACGCAC
IK25:pccB_rev 2013-07-16 Colony-PCR of methylmalonyl-CoA synthesis pathway GGTGATGAACATGTGGCTGGTCTG
IK26:BBa_I746200_fw 2013-07-19 Forward primer for permeability device ATGAACAAGAAGATTCATTCCCTGGCCTTG
IK27:BBa_I746200-BBa_B0029_rev 2013-07-19 Reverse primer for permeability device with partial Gibson overhang for BBa_B0029 RBS GAATACCAGT TCAGAAGTGGGTGTTTACGCTCATATAC
IK28:pccB-BBa_B0029-BBa_I746200_fw 2013-07-19 Forward primer for pccB with BBa_B0029 RBS and Gibson overhang for permeability device ATATGAGCGTAAACACCCACTTCTGAACTGGTATTCACACAGGAAACCTACTAG ATGTCCGAGCCGGAAGAGC
IK29:accA2-BBa_B0030_rev 2013-07-19 Reverse primer for accA2 with partial Gibson overhang for BBa_B0030 RBS TAATGAAGTTG CTAGTGATTCTCGCAGATGGC
IK30:sfp-BBa_B0030-accA2_fw 2013-07-19 Forward primer for sfp with BBa_B0030 RBS and Gibson overhang for accA2 TCCGGCGCCGCCATCTGCGAGAATCACTAGCAACTTCATTAAAGAGGAGAAATACTAG ATGAAGATTTACGGAATTTATATGGAC
IK31:sfp_rev 2013-07-19 Reverse primer for sfp
IK32:BBa_J04450-sfp_fw 2013-07-19 Forward primer for mRFP-containing backbones with Gibson overhang for sfp ATCACAATGGTCTCGTACGAAGAGCTTTTATAA TACTAGAGCCAGGCATCAAATAAAACG
IK33:BBa_J04450-BBa_I746200_rev 2013-07-19 Reverse primer for mRFP-containing backbones with Gibson overhang for permeability device ACAAGGCCAGGGAATGAATCTTCTTGTTCAT CTAGTATTTCTCCTCTTTCTCTAGTATG
IK34:pccB_fw 2013-07-19 Forward primer for pccB ATGTCCGAGCCGGAAGAGC
IK35:BBa_J04450-pccB_rev 2013-07-19 Reverse primer for mRFP-containing backbones with Gibson overhang for pccB ATGTCGGGCTGCTGCTCTTCCGGCTCGGACAT CTAGTATTTCTCCTCTTTCTCTAG
IK36:pLF03_seq_rev 2013-07-26 reverse primer for pLF03 for sequencing CCGGTATCAACAGGGACACCAG
IK37:pLF03-catR_seq_rev 2013-07-26 reverse primer for pLF03 sequencing CATTGAGCAACTGACTGAAATGCCTC
IK38:pIK1_fw 2013-08-30 Forward mutagenic primer for pIK1 ATTCCCTGGCCTTGT t GGTCAATCTGGGGATTTATG
IK39:pIK_rev 2013-08-30 Reverse mutagenic primer for pIK1 CCCAGATTGACC a ACAAGGCCAGGGAATGAATCTTC
IK40:pIK2-BBa_J23114-BBa_B0030_rev 2013-09-06 reverse primer for pIK2 with Gibson overhang for BBa_J23114-BBa_B0030 TTTAATCTCTAGAGCTAGCATTGTACCTAGGACTGAGCTAGCCATAAA CTCTAGTAGAGAGCGTTCAC
IK41:BBa_I746200-BBa_B0030-BBa_J23114_fw 2013-09-06 forward primer for BBa_I746200 with Gibson overhang for BBa_B0030-BBa_J23114 ATGCTAGCTCTAGAGATTAAAGAGGAGAAATACTAG ATGAACAAGAAGATTCATTCCCTG
IK42:BBa_I746200_rev 2013-09-06 reverse primer for BBa_I746200 TCAGAAGTGGGTGTTTACGCTC
IK43:pIK2-BBa_I746200_fw 2013-09-06 forward primer for pIK2 with Gibson overhang for BBa_I746200 TGAGCGTAAACACCCACTTCTGA TACTAGAGTCACACTGGCTC
NK_01_FS_62_screening_L_BB(without_mRFP)_rv2013-08-02 Primer for screening/sequencing of pFSN constructGTTCACCGACAAACAACAGATAAAACG
ygfG21C1 2013-06-03 For insertion of methylmalonyl-CoA synthesis pathway into E. coli. Primer from <bib id="pmid17959404"/>. TCACCGCGCACCGGCCTGCGGCAGCTCAGTGAATTTATCC AGATCTCGATCCC
ygfG21C2 2013-06-03 For insertion of methylmalonyl-CoA synthesis pathway into E. coli. Primer from <bib id="pmid17959404"/>. GTTATGCTGGATAATTTCTGCCGCTTCATTGGCGGTCATCC CAAAAAACCCCTCAAG

Module shuffling

Identifier Order date Note Sequence
AT01:RFC10prefix_TycA_fw 2013-08-12 Fw primer for amplification of TycAdCom; introduction of RFC10 prefix TTTT GAATTC GCGGCCGC T TCTAG ATG TTA GCA AAT CAa GCC AAT C
AT02:RFC10suffix_TycA_rv 2013-08-12 Rv primer for amplification of TycAdCom; introduction of RFC10 suffix TTTT CTGCAG CGGCCGC T ACTAGT A aGT TCG tTC TAC TTC TTT TTT C
AT03:RFC10prefix-TycB1_fw 2013-08-12 Fw primer for amplification of TycB1dCom; introduction of RFC10 prefix TTTT GAATTC GCGGCCGC T TCTAG ATG AGT GTA TTT AGC AAA GAA CAA G
AT04:RFC10suffix_TycB1_rv 2013-08-12 Rv primer for amplification of TycB1dCom; introduction of RFC10 suffix TTTT CTGCAG CGGCCGC T ACTAGT A TTC CTC CCC aCC TTC
AT05:RFC10prefix-TycC5_fw 2013-08-12 Fw primer for amplification of TycC5; introduction of RFC10 prefix TTTT GAATTC GCGGCCGC T TCTAG AG GCG CAT ATT GCa GAG AG
AT06:RFC10suffix_TycC5_rv 2013-08-14 Rv primer for amplification of TycC5; introduction of RFC10 suffix TTTT CTGCAG CGGCCGC T ACTAGT A TTT GGC TGT CTC TTC GAT GAA C
AT07:RFC10prefix-TycC6_fw 2013-08-12 Fw primer for amplification of TycC6; introduction of RFC10 prefix TTTT GAATTC GCGGCCGC T TCTAG AG GGG AAT GTC TTC TCG ATC
AT08:RFC10suffix_TycC6_rv 2013-08-14 Rv primer for amplification of TycC6; introduction of RFC10 suffix TTTT CTGCAG CGGCCGC T ACTAGT A TTA TTT CAG GAT aAA CAG TTC TTG
AT09:R10_B1_fw_longer 2013-08-18 Fw primer (longer) for amplification of TycB1dCom; introduction of RFC10 prefix TTTT GAA TTC GCG GCC GCT TCT AG ATG AGT GTA TTT AGC AAA GAA CAA GTT C
AT10:R10_B1_rv_longer 2013-08-18 Rv primer (longer for amplification of TycB1dCom; introduction of RFC10 suffix TTTT CTG CAG CGG CCG CTA CTA GTA ATA CGC aCT TTC CTC CCC GCC
AT11:R10_C5_fw_rpos 2013-08-18 Fw primer (repositioned) for amplification of TycC6; introduction of RFC10 prefix TTTT GAATTC GCGGCCGC T TCTAG AG GAGCAGTTCGAGACGATCCAGCC
AT101 2013-09-05 forward screening primer TycAdCom GGACATCATCGAACAGGC
AT102 2013-09-05 reverse screening primer TycAdCom GACATAGGCAAGATCCGTAG
AT103 2013-09-05 forward screening primer TycAdCom GCAAGGACAAGTAGATGGC
AT104 2013-09-05 reverse screening primer TycAdCom GTACAGCCATACAGTCGC
AT105 2013-09-05 forward screening primer TycC5 GTCACCATGCTGAGAACG
AT106 2013-09-05 reverse screening primer TycC5 CAGCAAAGCGGTCATAATC
AT107 2013-09-05 forward screening primer TycC6 AAGCTACGCTGTTGATTGC
AT108 2013-09-05 reverse screening primer TycC6 AGCACGTAACGATCCTC
IK09:TycA_A1_fw 2013-07-08 Colony-PCR of Brevibacillus parabrevis: TycA A domain ATGTTAGCAAATCAAGCCAATCTC
IK10:TycA_A1_rev 2013-07-08 Colony-PCR of Brevibacillus parabrevis: TycA A domain TTGGTTTGCTGTAAGATCAGGCTC
IK11:TycB_A1_fw 2013-07-08 Colony-PCR of Brevibacillus parabrevis: TycB A1 domain AATTCGGGAGTCGAGCTTTGTCAG
IK12:TycC6_rev 2013-07-19 reverse primer for tyrocidine TycC6 module TTATTTCAGGATGAACAGTTCTTGCAGG
IK13:IK13:TycB1-dCom-dC_fw 2013-07-19 forward primer for tyrocidine TycB1 module without Com and C domain GATTGCGTGGCAAACAATTCGGGAGTC
IK14:TycB1-TycC6_rev 2013-07-19 reverse primer for tyrocidine TycB1 module with Gibson overhang for TycC6 CAGGCTCGATCGAGAAGACATTCCC TTCCTCCCCGCCTTCCACATACGC
IK15:TycC6-TycB1_fw 2013-07-19 forward primer for tyrocidine TycC6 module with Gibson overhang for TycB1 GCGTATGTGGAAGGAGGGGAGGAA GGGAATGTCTTCTCGATCGAGCCTG
IK16:TycA_fw 2013-07-19 forward primer for tyrocidine TycA module ATGTTAGCAAATCAGGCCAATCTCATC
IK17:TycA-dCom-TycC5_rev 2013-07-19 reverse primer for tyrocidine TycA module without Com domain with Gibson overhang for TycC5 GAATGCGCTCTCGGCAATATGGGC TGTTCGCTCTACTTCTTTTTTCTCGG
IK18:TycC5-TycA-dCom_fw 2013-07-19 forward primer for tyrocidine TycC5 module with Gibson overhang for TycAdCom CCGAGAAAAAAGAAGTAGAGCGAACA GCCCATATTGCCGAGAGCGCATTC
IK19:TycB1-TycC5_rev 2013-07-19 reverse primer for tyrocidine TycB1 module with Gibson overhang for TycC5 GAATGCGCTCTCGGCAATATGGGC TTCCTCCCCGCCTTCCACATACGC
IK20:TycC5-TycB1_fw 2013-07-19 forward primer for tyrocidine TycC5 module with Gibson overhang for TycB1 GCGTATGTGGAAGGAGGGGAGGAA GCCCATATTGCCGAGAGCGCATTC
IK21:pSB4K5-TycB1dComdC_rev 2013-07-19 reverse primer for mRFP-carrying backbones with Gibson overhang for tyrocidine TycA module (nomenclature wrong) GTCGATGAGATTGGCCTGATTTGCTAACAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
IK22::pSB4K5-TycC6_fw 2013-07-19 forward primer for mRFP-carrying backbones with Gibson overhang for tyrocidine TycC6 module AACATCCTGCAAGAACTGTTCATCCTGAAA TAATAACGCTGATAGTGCTAGTGTAGATC
IK23:pSB4K5-TycA_rev 2013-07-19 reverse primer for mRFP-carrying backbones with Gibson overhang for tyrocidine TycB1 module without Com and C domains + start codon (nomenklature wrong) GACTCCCGAATTGTTTGCCACGCAATCCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
PW01:TycB_A1_rev 2013-07-08 Colony-PCR of Brevibacillus parabrevis: TycB A1 domain CTTGGCACTTCCTTCAGGCTTC
PW02:TycB_E1_fw 2013-07-08 Colony-PCR of Brevibacillus parabrevis: TycB E domain CGAGAGAGCGAGCAAGGTG
PW03:TycB_E1_rev 2013-07-08 Colony-PCR of Brevibacillus parabrevis: TycB E domain GTACTCGCCTTCTTCTTTTGC
PW04:pSB1C3-TycC5ΔC_rev 2013-07-19 Integration of Tetrapeptide NRPS from Brevibacillus parabrevis in backbone; Gibson primer for Tetrapeptide I & II TGTTTTGGTTGCGAGGAAGCTGTGCAGCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
PW05:TycC5ΔC_fwd 2013-07-19 Amplification of TycC5-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide I & II ATGCTGCACAGCTTCCTCGCAACCAAAACAGCC
PW06:TycC5ΔC-TycB1ΔCom_rev 2013-07-19 Amplification of TycC5-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide I & II GTGAAACAGCATCCCCTCTTGCATCGG AGGCTCGATCGAGAAGACATTCCCTTTG
PW07:TycC5ΔC-TycB1ΔCom_fwd 2013-07-19 Amplification of TycB1+C(TycB2)-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide I & II CAAAGGGAATGTCTTCTCGATCGAGCCT CCGATGCAAGAGGGGATGCTGTTTCAC
PW08:C(TycB2)-TycAΔCom_rev 2013-07-19 Amplification of TycB1+C(TycB2)-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide I & II GTTGTCGATGAGATTGGCCTGATTTGCTAACAT GATTTGCGCCAGCTCCTGCTCCGTGTT
PW09:C(TycB2)-TycAΔCom_fwd 2013-07-19 Amplification of TycA-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide I & II AACACGGAGCAGGAGCTGGCGCAAATC ATGTTAGCAAATCAGGCCAATCTCATCGACAAC
PW10:TycAΔCom-TycC6_rev 2013-07-19 Amplification of TycAΔCom-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide I CTTTGGCTGTCTCTTCGATGAACGC TCGCTCTACTTCTTTTTTCTCGGTGCAATG
PW11:TycAΔCom-TycC6_fwd 2013-07-19 Amplification of TycC6-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide I CATTGCACCGAGAAAAAAGAAGTAGAGCGA GCGTTCATCGAAGAGACAGCCAAAG
PW12:TycAΔE-TycC6_rev 2013-07-19 Amplification of TycAΔE-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide II; one mismatch C->t CTTTTGCACAGGCTCGATCGAGAAGAC GCTtTTGACAAAAAGAGCAACCTG
PW13:TycAΔE-TycC6_fwd 2013-07-19 Amplification of TycC6-Module from Brevibacillus parabrevis; Gibson primer for Tetrapeptide II; one mismatch G->a CAGGTTGCTCTTTTTGTCAAaAGC GTCTTCTCGATCGAGCCTGTGCAAAAG
PW14:C(TycC2)-indC_rev 2013-08-16 Amplification of C-domain from TycC2 from Brevibacillus parabrevis; Gibson overhang to IndC; for construct 1, 2 & 3 ACATTGTGTAATATTATTTTCTAACAT CGTTTTGCTGCTGGCAGGCTG
PW15:C(TycC2)-indC_fwd 2013-08-16 Amplification of indC from Photorhabdus luminescens; Gibson overhang to C-domain from TycC2; for construct 1, 2 & 3 CAGCCTGCCAGCAGCAAAACG ATGTTAGAAAATAATATTACACAATGT
PW16:indC_rev 2013-08-16 Amplification of indC from Photorhabdus luminescens; no Gibson overhang; for construct 1, 2 & 3 TTAGATTATTTTCTCAATCTCAGCAACACCTTC
PW17:TycAdE-C(TycC2)_rev 2013-08-16 Amplification of TycAdE from Brevibacillus parabrevis; Gibson overhang to C-domain from TycC2; for construct 1 CGAAAGGAAGCGGGCCAGCTC AGCAACCTGCTCGATCGTCGGGTA
PW18:TycAdE-C(TycC2)_fwd 2013-08-16 Amplification of C-domain from TycC2-module from Brevibacillus parabrevis; Gibson overhang to TycAdE; for construct 1 TACCCGACGATCGAGCAGGTTGCT GAGCTGGCCCGCTTCCTTTCG
PW19:TycC4dC_fwd 2013-08-16 Amplification of TycC4-module from Brevibacillus parabrevis without the C-domain; no Gibson overhang, ATG added; for construct 3 ATGTATCCGCGCGATCTGACGATTC
PW20:TycC4dC-C(TycC2)_rev 2013-08-16 Amplification of TycC4-module from Brevibacillus parabrevis without the C-domain; Gibson overhang to C-domain form TycC2; for construct 3 GGTGTACTCGGTTTTTTCCGA AATATGCGCAGCCAACTCATG
PW21:TycC4dC-C(TycC2)_fwd 2013-08-16 Amplification of C-domain from TycC2-module from Brevibacillus parabrevis; Gibson overhang to TycC4dC; for construct 3 CATGAGTTGGCTGCGCATATT TCGGAAAAAACCGAGTACACC
PW22:pSB1C3-TycC4dC_rev 2013-08-16 Insertion of construct 3 into pSB1C3-backbone, amplification of pSB1C3; Gibson overhang to TycC4dC; for construct 3 GAATCGTCAGATCGCGCGGATACAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
PW23:indC-pSB1C3_fwd 2013-08-16 Insertion of either fragments into pSB1C3-backbone, amplification of pSB1C3; Gibson overhang to indC from Photorhabdus luminescens; for constructs 1, 2 & 3 GGTGTTGCTGAGATTGAGAAAATAATCTAA TAATAACGCTGATAGTGCTAGTGTAGATC
PW24:TycC1dC_fwd 2013-08-16 Amplification of the TycC1-module from Brevibacillus parabrevis without the C-domain; no Gibson overhang, ATG added; for construct 2 ATGCAGACGAACAAACAACAGACG
PW25:pSB1C3-TycC1dC 2013-08-16 Insertion of construct 2 in pSB1C3-backbone, amplification of pSB1C3; Gibson overhang to TycC1-module without C-domain; for construct 2 CGTCTGTTGTTTGTTCGTCTGCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
PW26:TycC5dC_fwd 2013-09-04 Amplification of TycC5-module without C-domain from Brevibacillus parabrevis; no Gibson overhang, ATG added; for constructs A & B ATGCTGCACAGCTTCCTCGCAACC
PW27:TycC5dC-C(TycC4)_rev 2013-09-04 Amplification of TycC5-module without C-domain from Brevibacillus parabrevis; Gibson overhang to C-domain from TycC4; for constructs A & B CACATACGTCTCTTTTCCGCTCGT TTCGATGAACGCCGCCAGTTC
PW28:TycC5dC-C(TycC4)_fwd 2013-09-04 Amplification of C-domain from TycC4-module from Brevibacillus parabrevis; Gibson overhang to TycC5-module without C-domain; for constructs A & B GAACTGGCGGCGTTCATCGAA ACGAGCGGAAAAGAGACGTATGTG
PW29:C(TycC4)-TycC4dC_rev 2013-09-04 Amplification of C-domain from TycC4-module from Brevibacillus parabrevis; Gibson overhang to TycC4-module without C-domain; for constructs A, B, C, E & G GAATCGTCAGATCGCGCGGATA GGCAAACGTGTTGTTGAAATC
PW30:C(TycC4)-TycC4dC_fwd 2013-09-04 Amplification of TycC4-module without C-domain from Brevibacillus parabrevis; Gibson overhang to C-domain from TycC4-module; for constructs A, B, C, E & G GATTTCAACAACACGTTTGCC TATCCGCGCGATCTGACGATTC
PW31:TycC4-C(TycC4)_rev 2013-09-04 Amplification of TycC4-module from Brevibacillus parabrevis; Gibson overhang to C-domain from TycC4-module; for constructs B & C CACATACGTCTCTTTTCCGCTCGT GGCAATATGCGCAGCCAACTCATG
PW32:TycC4-C(TycC4)_fwd 2013-09-04 Amplification of C-domain from TycC4-module from Brevibacillus parabrevis; Gibson overhang to TycC4-module; for constructs B & C CATGAGTTGGCTGCGCATATTGCC ACGAGCGGAAAAGAGACGTATGTG
PW33:TycC6dTE-C(TycC2)_rev 2013-09-04 Amplification of TycC6-module without the TE-domain from Brevibacillus parabrevis; Gibson overhang to C-domain from TycC2-module; for constructs D & F GGTGTACTCGGTTTTTTCCGA CGTGATGAAATCGGCCACCTTTTC
PW34:TycC6dTE-C(TycC2)_fwd 2013-09-04 Amplification of C-domain from TycC2-module from Brevibacillus parabrevis; Gibson overhang to TycC6-module without TE-domain; for constructs D & F GAAAAGGTGGCCGATTTCATCACG TCGGAAAAAACCGAGTACACC
PW35:TycC6dTE-C(TycC4)_rev 2013-09-04 Amplification of TycC6-module without the TE-domain from Brevibacillus parabrevis; Gibson overhang to C-domain from TycC4-module; for constructs E & G CACATACGTCTCTTTTCCGCTCGT CGTGATGAAATCGGCCACCTTTTC
PW36:TycC6dTE-C(TycC4)_fwd 2013-09-04 Amplification of C-domain from TycC4-module from Brevibacillus parabrevis; Gibson overhang to TycC6-module without TE-domain; for constructs E & G GAAAAGGTGGCCGATTTCATCACG ACGAGCGGAAAAGAGACGTATGTG
PW37:pSB1C3-TycC5dC_rev 2013-09-04 Insertion of constructs A & B in pSB1C3-backbone, amplification of pSB1C3; Gibson overhang to TycC5-module without C-domain; for constructs A & B GGTTGCGAGGAAGCTGTGCAGCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG

Linker Variation

Identifier Order date Note Sequence
AR01 2013-09-17 sTycC4dC_fw ATG AAGCACTTGCTGCCGCTCGTC
AR02 2013-09-17 c(TycC2)s_rev TTGTGTAATATTATTTTCTAACAT CGCTACCTGTTGCAACCGCTC
AR03 2013-09-17 s-Ind_fw GAGCGGTTGCAACAGGTAGCG ATGTTAGAAAATAATATTACAATGT
AR04 2013-09-17 c(TycC2)l_rev_lTTGTGTAATATTATTTTCTAACAT AAGCGACTGCTGGTTCGCG
AR05 2013-09-17 l-Ind_fwAACGCGAACCAGCAGTCGCTT ATGTTAGAAAATAATATTACACAATGT
AR06 2013-09-17 lTycC4dC_fwATG CGCAACTATCCGGTCGAGACG
AR07 2013-09-17 TycC4dCom_short-BB_revGACGAGCGGCAGCAAGTGCTTCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
AR08 2013-09-17 TycC4dCom_long-BB_revCGTCTCGACCGGATAGTTGCGCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG
PW19:TycC4dC_fwd 2013-08-16 Amplification of TycC4-module from Brevibacillus parabrevis without the C-domain; no Gibson overhang, ATG added; for construct 3 ATGTATCCGCGCGATCTGACGATTC
PW20:TycC4dC-C(TycC2)_rev 2013-08-16 Amplification of TycC4-module from Brevibacillus parabrevis without the C-domain; Gibson overhang to C-domain form TycC2; for construct 3 GGTGTACTCGGTTTTTTCCGA AATATGCGCAGCCAACTCATG
PW21:TycC4dC-C(TycC2)_fwd 2013-08-16 Amplification of C-domain from TycC2-module from Brevibacillus parabrevis; Gibson overhang to TycC4dC; for construct 3 CATGAGTTGGCTGCGCATATT TCGGAAAAAACCGAGTACACC
PW14:C(TycC2)-indC_rev 2013-08-16 Amplification of C-domain from TycC2 from Brevibacillus parabrevis; Gibson overhang to IndC; for construct 1, 2 & 3 ACATTGTGTAATATTATTTTCTAACAT CGTTTTGCTGCTGGCAGGCTG

Domain shuffling and PPTases

Identifier Order date Note Sequence
KH1_ccdb_fw 2013-07-23 amplifing ccdb with RBS from pDONR TGGTGCCATTACATACAGATACT GAGAACAGGGGCTGGTGAAATGC
KH2_ccdb_rv 2013-07-23 amplifing ccdb with RBS from pDONR AGAGTCTGTCTGTTCAATCCACTT TTATATTCCCCAGAACATCAGGTTAATGGCG
KH3_indC_backbone-withoutT_fw 2013-07-23 linearization of vector with partial indigoidine synthase AAGTGGATTGAACAGACAGACTCTAAAAC
KH4_indC_backbone-withoutT_rv 2013-07-23 linearization of vector with partial indigoidine synthase AGTATCTGTATGTAATGGCACCAATAGACGC
KH5_indC_T_fw 2013-07-23 introducing T domain of indC TGGTGCCATTACATACAGATACT GAAATAAGGCTTGGAAAAATTTGGATGGAAGT
KH6_indC_T_rv 2013-07-23 introducing T domain of indC AGAGTCTGTCTGTTCAATCCACTT AGCCAATTCTGCTATATTAGGAGATTGA
KH7_bpsA_T_fw 2013-07-23 introducing T domain of bpsA TGGTGCCATTACATACAGATACT GAGAAGGAAATCGCAGCCGTGTGGG
KH8_bpsA_T_rv 2013-07-23 introducing T domain of bpsA AGAGTCTGTCTGTTCAATCCACTT GGCCAGCTTTTCAATTGTTGGTGACTCC
KH9_ccdB-Big_fw 2013-08-06 ccdB_Big amplified from pDONOR (Dominik) for indC(ccdB) construct TGCCATTACATACAGATACT ACTGGCTGTGTATAAGGGAGCCTGAC
KH10_ccdB-Big_rv 2013-08-06 ccdB_Big amplified from pDONOR (Dominik) for indC(ccdB) construct AGAGTCTGTCTGTTCAATCCACTT CGCGTGGATCCGGCTTAC
KH11_plu2642-T_fw 2013-08-14 T domain exchange plu2642 TGGTGCCATTACATACAGATACT GAAACACAGATCGTAAAGATATGG
KH12_plu2642-T_rv 2013-08-14 T domain exchange plu2642 AGAGTCTGTCTGTTCAATCCACTT TGCCAATTGTTTAACCGTTG
KH13_BBa-entD_fw 2013-08-09 submitting endD to registry GCAT GAATTCGCGGCCGCTTCTAG ATGAAAACTACGCATACCTCCCTC
KH14_BBa-entD_rv 2013-08-09 submitting endD to registry GCAT CTGCAGCGGCCGCTACTAGTA TCATTAATCGTGTTGGCACAGCG
KH15_BBa-sfp_Bsub_fw 2013-08-09 submitting sfp_Bsub to registry GCAT GAATTCGCGGCCGCTTCTAG ATGAAGATTTACGGAATTTATATGGAC
KH16_BBa-sfp_Bsub_rv 2013-08-09 submitting sfp_Bsub to registry GCAT CTGCAGCGGCCGCTACTAGTA TCATTATAAAAGCTCTTCGTACGAGAC
KH17_BBa-svp_Svert_fw 2013-08-09 submitting svp_Svert to registry needed template: pRB04 GCAT GAATTCGCGGCCGCTTCTAG A TGATCGCCGCCCTCCTGCCCTCC
KH18_BBa-svp_Svert_rv 2013-08-09 submitting svp_Svert to registry needed template: pRB04 GCAT CTGCAGCGGCCGCTACTAGTA AGCTAGCTCATTACGGGAC
KH19_BBa-delC_fw 2013-08-09 submitting delC to registry GCAT GAATTCGCGGCCGCTTCTAG ATGCAGCTCGTGTCCGTGCG
KH20_BBa-delC_rv 2013-08-09 submitting delC to registry GCAT CTGCAGCGGCCGCTACTAGTA TTA TCATGTCGATTCCTTGGTGC
KH21_BBa-indC_fw 2013-08-09 submitting indC to registry GCAT GAATTCGCGGCCGCTTCTAG ATGTTAGAAAATAATATTACACAATG
KH22_BBa-indC_rv 2013-08-09 submitting indC to registry GCAT CTGCAGCGGCCGCTACTAGTA TTAGATTATTTTCTCAATCTCAG
NI01:bpsA_AOxA_PstI_fw 2013-06-04 for amplifying bpsA (AOxA domain) GAGGAGAAATACTAGATGACACTGCAGGAAACAAGCGTGC
NI02:bpsA_AOxA_rv 2013-06-04 for amplifying bpsA (AOxA domain) GAAGGGCCGTTCCACCAGCTCAGCGTTGACCTGGTCAGAAGCG
NI03:bpsA_T_fw 2013-06-04 for amplifying bpsA (T domain) GACCAGGTCAACGCTGAGCTGGTGGAACGGCCCTTCGTCG
NI04:bpsA_T_rv 2013-06-04 for amplifying bpsA (T domain) GACGAAGCGACTAGACTCCTGAGCCACTTCTCTCTCCAGCCG
NI05:bpsA_TE_fw 2013-06-04 for amplifying bpsA (TE domain) with RBS 1 GAGAGAGAAGTGGCTCAGGAGTCTAGTCGCTTCGTCCGA
NI06:bpsA_TE_Stop_XbaI_rbs_rv 2013-06-04 for amplifying bpsA (TE domain) with RBS 1 TGGCAGCAGAGCAGCGATCATCTAGTATTTCTCCTCTTTCCTCTAGATCATCATTCCCCCAGCAGGTATCTAAT
NI07:svp_fw 2013-06-04 for amplifying svp GAGGAGAAATACTAGATGATCGCTGCTCTGCTGCCAAGTTGG
NI08:svp_rv 2013-06-04 for amplifying svp GCACTATCAGCGTTATTATGGCACGGCAGTCCTATCGTCG
NI09:pSB1C3_fw 2013-06-04 for linearizing, amplifying pSB1C3 GATAGGACTGCCGTGCCATAATAACGCTGATAGTGCTAGTGTAGATCGC
NI10:pSB1C3_PstI_rv 2013-06-04 for linearizing, amplifying pSB1C3 GCTTGTTTCCTGCAGTGTCATCTAGTATTTCTCCTCTTTCTCTAGTATGTG
NI11:bpsA_rvN 2013-07-15 for amplifying bpsA (TE domain) with RBS 2 TGGCAGCAGAGCAGCGATCATTATTTAGGTTTCCTGTGTGAATCATCATTCCCCCAGCAGGTATCTAAT
NI12:svp_fwN 2013-07-15 for amplifying svp with RBS 2 CAGGAAACCTAAATAATGATCGCTGCTCTGCTGCCAAGTTGG
NI13_ccdb_fw 2013-07-23 amplifing ccdb with RBS from pDONR CGTCGCACCTAGGACCGAAACA GAGAACAGGGGCTGGTGAAATGC
NI14_ccdb_rv 2013-07-23 amplifing ccdb with RBS from pDONR GCCACTTCTCTCTCCAGCCGTCG TTATATTCCCCAGAACATCAGGTTAATGGCG
NI15_bpsA_backbone-withoutT_fw 2013-07-23 linearization of vector with partial indigoidine synthase CGACGGCTGGAGAGAGAAGTGGC
NI16_bpsA_backbone-withoutT_rv 2013-07-23 linearization of vector with partial indigoidine synthase TC TGTTTCGGTCCTAGGTGCGACG
NI17_indC_T_fw 2013-07-23 introducing T domain of indC CGTCGCACCTAGGACCGAAACA GAAATAAGGCTTGGAAAAATTTGGATGGAAGT
NI18_indC_T_rv 2013-07-23 introducing T domain of indC GCCACTTCTCTCTCCAGCCGTCG AGCCAATTCTGCTATATTAGGAGATTGA
NI19_bpsA_T_fw 2013-07-23 introducing T domain of bpsA CGTCGCACCTAGGACCGAAACA GAGAAGGAAATCGCAGCCGTGTGGG
NI20_bpsA_T_rv 2013-07-23 introducing T domain of bpsA GCCACTTCTCTCTCCAGCCGTCG GGCCAGCTTTTCAATTGTTGGTGACTCC
NI19:ngrA_fw GAATTCGCGGCCGCTTCTAG ATGGAACAGACAGTTATACACACC
NI20:ngrA_rv CTGCAGCGGCCGCTACTAGTA TTATTGAATAATTAGCGTTAATATTTCCTG
RB05:RBS1-BpsA_fw2013-07-01Gibson assembly of pSB1C3 bpsA svpAAAGAGGAGAAA TACTAG ATGACACTTCAGGAAACCAGCG
RB06:BpsA-RBS2_rv2013-07-01Gibson assembly of pSB1C3 bpsA svpCTTTCCTGTGTGA AAGCTT TCA TCACTCCCCGAGCAGATATCG
RB07:RBS2-svp_fw2013-07-01Gibson assembly of pSB1C3 bpsA svpCTTTCACACAGGAAAG TAAATA ATGGCTGCTCTTCTTCCTAGTTGGGC
RB08:svp-pSB1C3_rv2013-07-01Gibson assembly of pSB1C3 bpsA svpCTATCAGCGTTATTA AAGCTT TCATCA TGGCACGGCAGTCCTATCG
RB09:pSB1C3_fw2013-07-01Gibson assembly of pSB1C3 bpsA svpAAGCTT TAATAACGCTGATAGTGCTAGTGTAGATCGC
RB10:pSB1C3-RBS1_rv2013-07-01Gibson assembly of pSB1C3 bpsA svpCTAGTA TTTCTCCTCTTT CTCTAGTATGTGTG
RB11:BpsA_Taka_fw2013-07-08colony PCR bpsA Takahashicat atgactcttcaggagaccagcgtgctc
RB12:BpsA_Taka_rv2013-07-08colony PCR bpsA Takahashiaag cttctcgccgagcaggtagcggatgtg
RB13:svp_Taka_fw2013-07-08colony PCR svp Takahashicat atgatcgccgccctcctgccctcctg
RB14:svp_Taka_rv2013-07-08colony PCR svp Takahashictcgagcgggacggcggtccggtcgtccgc
RB15:svp_Sanch_fw2013-07-08PCR svp SanchezTATA ATGCATGCTCGCCGCCCTCCCC
RB16:svp_Sanch_rv2013-07-08PCR svp SanchezTTAAGATCTCGGGACGGCGGTCCGGTC
RB17:sfp_fw2013-07-08sfp extraction from BAP1ATGAAGATTTACGGAATTTATATGG
RB18:sfp_rv2013-07-08sfp extraction from BAP1TTATAAAAGCTCTTCGTACGAGACC
RB19:entD_fw2013-07-08entD extraction from E.coli LambalotTAAATA ATGGTCGATATGAAAACTACGC
RB20:entD_rv2013-07-08entD extraction from E.coli LambalotAAGCTT ATTAATCGTGTTGGCACAGCG
RB21:pSB1C3_fw 2013-07-15 indigoidine exchangeable construct TAATGA GCTAGC TAATAACGCTGATAGTGCTAGTG
RB22:pSB1C3_rv 2013-07-15 indigoidine exchangeable construct CAT GGTACC TTTCTCCTCTTT CTCTAGTATGTGTG
RB23:bpsA-fus_fw 2013-07-15 indigoidine exchangeable construct CACACATACTAGAG AAAGAGGAGAAA GGTACC ATGACTAGTACACTGCAGG
RB24:bpsA-fus_rv 2013-07-15 indigoidine exchangeable construct CAT GGATCC GGTTTCCTGTGTGAA TCATTA TTCCCCCAGCAGGTATCTAATATG
RB25:svp-fus_fw 2013-07-15 indigoidine exchangeable construct CACAGGAAACC GGATCC ATGACTAGTATCGCTGCTCTGCTG
RB26:svp-fus_rv 2013-07-15 indigoidine exchangeable construct CAGCGTTATTA GCTAGC TCATTA TGGCACGGCAGTCCTATC
RB27:indC-Plum_fw 2013-07-15 indigoidine exchangeable construct CATACTAGAG AAAGAGGAGAAA GGTACC ATGTTAGAAAATAATATTACACAATG
RB28:indC-Plum_rv 2013-07-15 indigoidine exchangeable construct CAT GGATCC GGTTTCCTGTGTGAA TTA TTAGATTATTTTCTCAATCTCAG
RB29:svp-Svert_fw 2013-07-15 indigoidine exchangeable construct CACAGGAAACC GGATCC A TGATCGCCGCCCTC
RB30:svp-Svert_rv 2013-07-15 indigoidine exchangeable construct CAGCGTTATTA GCTAGC TCA TTACGGGACGGCGGTC
RB31:Sc-indC-Schr_fw 2013-07-15 indigoidine exchangeable construct GAG AAAGAGGAGAAA GGTACC ATGAGCGTAGAGACCATC
RB32:Sc-indC-Schr_rv 2013-07-15 indigoidine exchangeable construct CAT AGATCT GGTTTCCTGTGTGAA TTA TCAGTAGTTGGGCGTCTTG
RB33:entD-MG1655_fw 2013-07-15 indigoidine exchangeable construct CACAGGAAACC GGATCC ATGAAAACTACGCATACCTC
RB34:entD-MG1655_rv 2013-07-15 indigoidine exchangeable construct CAGCGTTATTA GCTAGC TCA TTAATCGTGTTGGCACAGC
RB35:sfp-Naka_fw 2013-07-15 indigoidine exchangeable construct CACAGGAAACC GGATCC ATGAAGATTTACGGAATTTATATGGAC
RB36:sfp-Naka_rv 2013-07-15 indigoidine exchangeable construct CAGCGTTATTA GCTAGC TCA TTATAAAAGCTCTTCGTACGAGAC
RB37:Plum-extr_fw 2013-07-15 genomic extraction ATGTTAGAAAATAATATTACACAATG
RB38:Plum-extr_rv
 2013-07-15 genomic extraction TTAGATTATTTTCTCAATCTCAG
RB39:Svert-extr_fw 2013-07-15 genomic extraction GTGATCGCCGCCCTC
RB40:Svert-extr_rv 2013-07-15 genomic extraction TTACGGGACGGCGGTC
RB41:MG1655-extr_fw 2013-07-15 genomic extraction ATGAAAACTACGCATACCTCCCTC
RB42:MG1655-extr_rv 2013-07-15 genomic extraction TTAATCGTGTTGGCACAGCGTTATG
RB43:Bsub-extr_fw 2013-07-15 genomic extraction ATGAAGATTTACGGAATTTATATGGAC
RB44:Bsub-extr_rv 2013-07-15 genomic extraction TTATAAAAGCTCTTCGTACGAGAC
RB45:Sc-indC-Schr_rvG 2013-07-15 indigoidine exchangeable construct CAT GGATCC GGTTTCCTGTGTGAA TTA TCAGTAGTTGGGCGTCTTG
RB46:indC-woPPT_rv 2013-08-05 indC-pSB1C3 w/o PPTase; RB21 overlap GCACTATCAGCGTTATTA GCTAGCTCATTA TTA TTAGATTATTTTCTCAATCTCAG
RB47:indC-SpeI_fw 2013-08-05 remove cutting sites from indC-Plum CAAGTTCCTAAACCC ACTAGT CTGGCTTAT ATTATTTATACCTCTGGTAGCAC
RB48:indC-EcoRI_rv 2013-08-05 remove cutting sites from indC-Plum CAATACCCACC GAATTC TTTGAGCT AATTTCTGACAGACAATACC
RB49:indC-EcoRI_fw 2013-08-05 remove cutting sites from indC-Plum AGCTCAAA GAATTC GGTGGGTATTG GGCTTTTTTGTGATC
RB50:indC-SpeI_rv 2013-08-05 remove cutting sites from indC-Plum ATAAGCCAG ACTAGT GGGTTTAGGAACTTG GAACTTGAACTGTG
RB51:pSB(2/3)K3_fw 2013-08-05 PPTase plasmid TAATGA GCTAGC tactagtagcggccgctgcagtc
RB52:pSB(2/3)K3_rv 2013-08-05 PPTase plasmid CAT GGATCC GGTTTCCTGTGTGAA ctctagaagcggccgcgaattcc
RB53:entF-T_fw 2013-08-07 bring T-Domain in indC TGGTGCCATTACATACAGATACT GAAACGATTATCGCCGCGGCATTC
RB54:entF-T_rv 2013-08-07 bring T-Domain in indC AGAGTCTGTCTGTTCAATCCACTT TGCCAGTTTGGCGACAGTTGACG
RB55:tycA1-T_fw 2013-08-07 bring T-Domain in indC TGGTGCCATTACATACAGATACT GAATCGATTCTCGTCTCCATCTGG
RB56:tycA1-T_rv 2013-08-07 bring T-Domain in indC AGAGTCTGTCTGTTCAATCCACTT AGCAACCTGCTCGATCGTCGGGTAATTC
RB57:tycC6-T_fw 2013-08-07 bring T-Domain in indC TGGTGCCATTACATACAGATACT GAACAGCAACTGGCAGCCATCTGGCAAG
RB58:tycC6-T_rv 2013-08-07 bring T-Domain in indC AGAGTCTGTCTGTTCAATCCACTT GGCCACCTTTTCGATCGTCGGATACTG
RB59:delH4-T_fw 2013-08-07 bring T-Domain in indC TGGTGCCATTACATACAGATACT GAGACGCTGCTGGCCCGTATCTG
RB60:delH4-T_rv 2013-08-07 bring T-Domain in indC AGAGTCTGTCTGTTCAATCCACTT CGCCAGCTCGGCAATGCTTTGC
RB61:delH5-T_fw 2013-08-07 bring T-Domain in indC TGGTGCCATTACATACAGATACT GCCATGGCGCTGGCCCGCATC
RB62:delH5-T_rv 2013-08-07 bring T-Domain in indC AGAGTCTGTCTGTTCAATCCACTT CAGCAGGCCGGCAATGGTCG
RB63:pSB(2/3)K3_rv_korr 2013-08-07 PPTase plasmid CAT GGATCC GGTTTCCTGTGTGAA CTCTAGTATGTGTGAAATTGTTATCC
RB64:DelC-extr_fw 2013-08-07 get Delftia PPTase ATGCAGCTCGTGTCCGTGCGTGAG
RB65:DelC-extr_rv 2013-08-07 get Delftia PPTase TCATGTCGATTCCTTGGTGCGCCAC
RB66:DelC-pSB2K3_fw 2013-08-07 DelC in PPTase-construct CACAGGAAACC GGATCC ATGCAGCTCGTGTCCGTGCGTGAG
RB67:DelC-pSB2K3_rv 2013-08-07 DelC in PPTase-construct CAGCGTTATTA GCTAGC TCA TCATGTCGATTCCTTGGTGCGCCAC
RB68:indC-SpeI_fw_corr 2013-08-09 indC w/o cutting sites corrected CAAGTTCCTAAACCC ACgAGT CTGGCTTAT ATTATTTATACCTCTGGTAGCAC
RB69:indC-EcoRI_rv_corr 2013-08-09 indC w/o cutting sites corrected CAATACCCACC GAAcTC TTTGAGCT AATTTCTGACAGACAATACC
RB70:indC-EcoRI_fw_corr 2013-08-09 indC w/o cutting sites corrected AGCTCAAA GAgTTC GGTGGGTATTG GGCTTTTTTGTGATC
RB71:indC-SpeI_rv_corr 2013-08-09 indC w/o cutting sites corrected ATAAGCCAG ACTcGT GGGTTTAGGAACTTG GAACTTGAACTGTG
RB72:bpsA-TTE_rv 2013-08-09 exchange T and TE-domain GCACTATCAGCGTTATTA GCTAGC TCATTA TTCCCCCAGCAGGTATCTAATATG
RB73:entF-TTE_rv 2013-08-09 exchange T and TE-domain GCACTATCAGCGTTATTA GCTAGC TCA TTACCTGTTTAGCGTTGCGCGAATAATCG
RB74:tycC6-TTE_rv 2013-08-09 exchange T and TE-domain GCACTATCAGCGTTATTA GCTAGC TCA TTATTTCAGGATGAACAGTTCTTGC
RB75:delH5-TTE_rv 2013-08-09 exchange T and TE-domain GCACTATCAGCGTTATTA GCTAGC TCATTA TCAGTCCAGCGCGTACTCCAG
RB76:plu2670-T_fw 2013-08-15 bring T-Domain in indC TGGTGCCATTACATACAGATACT GAAACCACACTGGCTGCTATC
RB77:plu2670-T_rv 2013-08-15 bring T-Domain in indC AGAGTCTGTCTGTTCAATCCACTT CGCAAATGCGGATAACACC
RB78:indC-SpeI_rv_corr2 2013-08-26 indC w/o cutting sites w/o frameshift ATAAGCCAG ACTcGT GGGTTTAGGAACTTG AACTGTG
RB79:indC_woPPTase_rv2 2013-08-29 indC-pSB1C3 w/o PPTase; RB35 5' part CAGCGTTATTA GCTAGC TCA TTAGATTATTTTCTCAATCTCAG
RB80:indCdT_3_fw 2013-09-06 T-domain border combinations bpsA GGCGGATATCCTATGAGTTTGAGATTGC
RB81:indCdT_3_rv 2013-09-06 T-domain border combinations bpsA TCCATTGGCCGTCAAAGGTAATTTATCG
RB82:bpsA-T3_fw 2013-09-06 T-domain border combinations bpsA CGATAAATTACCTTTGACGGCCAATGGA AAGATCGATGTGAAAGCACTGGCCGCTTCTGACC
RB83:bpsA-T3_rv 2013-09-06 T-domain border combinations bpsA GCAATCTCAAACTCATAGGATATCCGCC CAGACCTGGCCAGCAGATCACAGG
RB84:indCdT_4_fw 2013-09-06 T-domain border combinations bpsA TTTGAAGTTGCATACCAGCTTGAACAAGC
RB85:bpsA-T4_rv 2013-09-06 T-domain border combinations bpsA GCTTGTTCAAGCTGGTATGCAACTTCAAA TGCCACGCGAGCTCCGAAGCTATATCC
RB86:indC-T2_fw 2013-09-06 T-domain border combinations bpsA TTACAGATACTTTTTCAATCTCCTAATATAGCAG
RB87:indC-T2_rv 2013-09-06 T-domain border combinations bpsA TACTTCCATCCAAATTTTTCCAAGC
RB88:bpsA-T2_fw 2013-09-06 T-domain border combinations bpsA GCTTGGAAAAATTTGGATGGAAGTA CTGAGACGCGAAAATGCTAGTGTCC
RB89:bpsA-T2_rv 2013-09-06 T-domain border combinations bpsA TTAGGAGATTGAAAAAGTATCTGTAA AGGCAGGGACACTCCCAGTCTAGCATTCAG
RB90:indC-T-screen_fw 2013-09-13 screeening primer GATCAATGCGGCCTTTAATATTCG
RB91:bpsA-T-screen_fw 2013-09-13 screeening primer GGAACTGAATGCTAGACTGGGAG
RB92:entF-T3_fw 2013-09-16 native T with border combination 31 CGATAAATTACCTTTGACGGCCAATGGA AAGCTGGATCGCAAAGCCTTACCGTTG
RB93:tycA1-T3_fw 2013-09-16 native T with border combination 31 CGATAAATTACCTTTGACGGCCAATGGA AAGATCGACCGCAAAGCGTTGC
RB94:tycC6-T3_fw 2013-09-16 native T with border combination 31 CGATAAATTACCTTTGACGGCCAATGGA AAAGTGGATCGCAAAGCTTTG
RB95:delH4-T3_fw 2013-09-16 native T with border combination 31 CGATAAATTACCTTTGACGGCCAATGGA AAGCTGGACCGGCAGGCCCTG
RB96:delH5-T3_fw 2013-09-16 native T with border combination 31 CGATAAATTACCTTTGACGGCCAATGGA AAGCTTGACCGTGGTGCGCTG
RB97:plu2642-T3_fw 2013-09-16 native T with border combination 31 CGATAAATTACCTTTGACGGCCAATGGA AAAATCGATTTCGACACATTACAAG
RB98:plu2670-T3_fw 2013-09-16 native T with border combination 31 CGATAAATTACCTTTGACGGCCAATGGA AAGCTGGACCGTCGGGCGTTACCGGCAC
RB99:plu2642_pRB23-plu_fw 2013-09-24 plu2642 on pSB1C3 CATACTAGAG AAAGAGGAGAAA GGTACC ATGCAATCAACTCTCCCAATAATAAAATGG
RB100:plu2642_pRB23-plu_rv 2013-09-24 plu2642 on pSB1C3 CAGCGTTATTA GCTAGC TCA TCAACTCAAGAATCGAGCTAATTCGTTAACAC
RB101:plu2642_A_rv 2013-09-24 shuffling of all domains GTTCTGGCATCCTTCTTAATTAACAT TGCAGCGTGTTTCACTTGATCGATAAC
RB102:indC_Ox_fw 2013-09-24 shuffling of all domains ATGTTAATTAAGAAGGATGCCAGAAC
RB103:plu2642_valInd_fw 2013-09-24 shuffling of all domains CAGCCTGCCAGCAGCAAAACG ATGCAATCAACTCTCCCAATAATAAAATGG
RB104:plu2642_valInd_rv 2013-09-24 shuffling of all domains CAT CGTTTTGCTGCTGGCAGGCTG
RB105:tycC2_T3_fw 2013-09-24 shuffling of all domains CGATAAATTACCTTTGACGGCCAATGGA AAAGTGGATCGCAAGGCATTG
RB106:tycC2_T1_rv 2013-09-24 shuffling of all domains AGAGTCTGTCTGTTCAATCCACTT GGCCAGGCCCGCGATCGTTGG
RB107:tycC2_A_fw 2013-09-24 shuffling of all domains CATACTAGAG AAAGAGGAGAAA GGTACC ATGACGGAAGCGGAAAAACGCACACTCCT
RB108:tycC2_A_rv 2013-09-24 shuffling of all domains GTTCTGGCATCCTTCTTAATTAACAT GACGACCGCTTTGACCGCTTCATG
RB109:tycC2_valInd_fw 2013-09-24 shuffling of all domains CAGCCTGCCAGCAGCAAAACG ATGACGGAAGCGGAAAAACGCACACTCC
RB110:plu2642_TE_fw shuffling of all domains AGTGGATTGAACAGACAGACTCT AAAATCGTTGAAGGTGAAGTAACCAG

Table 7: Instruments

Instrument Type Manufacturer
Table Centrifuge Microfuge® 18 Centrifuge Beckman CoulterTM
Table Centrifuge Microfuge® 22R Centrifuge Beckman CoulterTM
Centrifuge Allegra X-12R Beckman CoulterTM
Shaker VORTEXGENIE 2 Scientific Industries, SITM
Heating plate (magnet stirrer)MR Hei-Standard Heidolph
Heatblock QBD4 Grant
Heatblock (shakeing function) Thermomixer comfort Eppendorf
PCR-machineMyCyclerTM thermo cyclerBioRad
PCR-machineT100 Therma CyclerBioRad
UV-chamber Transluminator Vilber Lourmat
Scale (fine)PioneerTM PA114C OHAUS
ScalePioneerTM PA4101C OHAUS
Fridge KTP 1750 Premium Liebherr
Freezer GP 1366 Premium Liebherr
HoodTischabzugWesemann® Laboreinrichtung
Draw-off pumpVacuhand controlVacubrand
Incubator HT Multitron Version 2 INFORS
Plate reader
Computer Sun Microsystems
UV/VIS Spectrometer Ultrospec 3300 pro Amersham Biosciences
Photometer NanoDrop® ND-1000 Spectrophotometer peQLab Biotechnologie GmbH
Photometer NanoVue General Electric
Ice Machine MF22SCOTSMAN®
Gel electrophoresis chamber Mupid®-One Advance
Cell density meter Ultrospec10 Amersham Biosciences
pH-Meter pH-Meter 765 Calimatic Knick
Incubator Heraeus Thermo Scientific
Lyophilizer Gefriertrocknungsanlage ALPHA 1-2 LD Plus Best. Nr. 101521 Christ
Ultra sonification stick Sonoplus Gm 2070 (2002)Bandelin
Vacuum Manifold Qiavac 24 Plus QIAGEN
Gaskartusche Ventil CV470 Burner 13883
NuPAGE(R) Novex 3-8% Tris-Acetate Gel 1 SDS-Gel Life Technologies GmbH

Table 8: Mass spectrometry at neonate screening

Instrument Type Manufacturer Order number
micro filter plates Multiscreen 96 well Merck Millipore MSHVN4550
microtiter plates 96 well Mikrotiter plates made of Polypropylen (F-Form) Greiner 655201
microtiter plates locks Micromats
aluminium foil
Mass spectrometer Quattro Ultima (ESI-MS/MS) Micromass (today: Waters)
Methanol (20837.320 METHANOL HIPERSOLV HPLC ISOCRATIC GRADE)

Solution 1 (for MS sample preparation): Neo Gen stable isotope standard kit A und B (NSK-AB, EurIsotop)
1. One vial of amino acid standard (NSK-A) is taken up in 1 ml methanol/water (1:1 v/v) and solubilized for ca. 15 minutes in an ultra-sonification bath. Caution: make sure that the lid is closed well and not wetted by water of the ultra-sonification bath.
2. One vial of acylcarnitine standard (NSK-B) is taken up in 1ml of methanol and solubilized for ca. 15 minutes in the ultra-sonification bath.Caution: make sure that the lid is closed well and not wetted by water of the ultra-sonification bath.
Contents of both vials are transfered quantitatively with methanol in a 200 ml graduate flask. The graduate flask is filled up to the calibration mark with methanol. Standard solutions get a consecutive number. When two 200 ml graduate flasks are prepared simultaneously, contents of both flasks can be combined, if levels of both standards (old and new) coincide. Afterwards the whole solution will be labeled with one common number.
Stability of solution 1: 7 days

Solution 2 (for MS sample preparation): butanolic hydrochloric acid (3 N), water-free:
1. 900 ml butanol (p.A., Merck 1.01990.1000) are cooled in an ice bath at 0°C (magnetic stirrer). The vessel has to be closed to avoid ice condensation of water.
2. 100 ml acetylchloride (p.A., Merck 1.00031.0250) are added dropwiese from a dropping funnel over a a period of 5 minutes while stirring constantly (protection goggles and gloves!).
Caution: make sure that no water or ice gets into the solution.
3. The solution is stirred for additional 10 minutes with closed lid on ice. Stability of solution 2: 1 year

Solution 1 (for MS sample preparation):
Acetonitrile p.A./water (1:1 v/v) (83639.320DE ACETONITRIL HIPERSOLV SUPER GR REAG.PE)
Stability of solution 3: 1 year

Table 9: Lab materials

Material Name Manufacturer
Bottle 50 ml, 100 ml, 200 ml, 500 ml, 1000 ml
Beaker 50 ml, 100 ml, 200 ml, 500 ml 1000 ml
Conical flask 500 ml, 1000 ml, 100 ml
Conical flask 300 ml
Single channel pipet Pipetman® P2, P20, P200, P1000 Gilson
Multichannel pipet
Multistep pipet
Pipet
0.2 ml PCR Tube, Flat Cap, natural I1402-8200 Starlab GmbH
Microcentrifuge tubes
Centrifugetubes
PCR 8-strips
Gloves Nitril Gr. L Starlab puderfr Starlab
Gloves Nitril Gr. M Starlab puderfr Starlab
Gloves Nitril Gr. S Starlab puderfr Starlab
Filtertips
Inoculating loop
Disposal bags
Flasks
Plate
PETRI DISHES P5731-500EA Sigma-Aldrich Chemie GmbH
PETRI DISHES N221.2 Carl Roth GmbH & Co.KG
TipOne® Pipette Tip S1110-1700 Starlab GmbH
TipOne® Pipette Tip, 1000μl, Graduated S1111-2721 Starlab GmbH
neoBox-81 6er Set, je 1 x transparent, g 22916
neoLab-Klebepunkte für Reaktionsgefäße 19079
Gene Pulser/MicroPulser Cuvettes, 0.1 cm 165-2089
neoLab-Papierschere, 23 cm lang, gerade 23272
TipOne® Pipette Tip, 200μl S1110-1700 Starlab GmbH
TipOne® Pipette Tip, 10μl, Graduated, Re S1111-3700 Starlab GmbH
PIPETTE, 5 ML 606180 Greiner bio-one GmbH
Bleiring mit Vinylbeschichtung, 57 mm In 310161013 NEOLAB GmbH
TipOne® Pipette Tip 10μl, Graduated, Rac S1111-3800 Starlab GmbH
Reaktionsgefäß,S.L.1,5ml,farbl. 12682 Eppendorf, Fisher Scientific GmbH
Reaktionsgefäß,S.L.,2ML, colorless 12776 Eppendorf
neoTape-Beschriftungsband, 13 mm, grau 280126114 NEOLAB GMBH
neoTape-Beschriftungsband, 25 mm, lachs 280126229 NEOLAB GMBH
10ml Serological Pipette, Filter, Steril E4860-1011 Starlab GmbH
Gloves Latex + Alovera L 14089
Gloves Latex + Alovera M 14088
Gloves Latex + Alovera S 14087
PH-stripes WHAT10362000 PANPEHA PLUS
100 Run24Barcode 20110099-100 GATC Biotech AG
Röhrchen konisch, Polypropylen, 50 ml 352070 NEOLAB GMBH
Gene Pulser/MicroPulser Cuvettes, 0.1 cm 165-2089 Bio-Rad Laboratories GmbH
0.2 ml 8-Tube Strips Without Caps, natur TBS-0201 Bio-Rad Laboratories GmbH
Rundbodenröhrchen, Polypropylen, 14 ml 352059 NEOLAB GMBH
Optical Flat 8-Cap Strips TCS-0803 Bio-Rad Laboratories GmbH
Impfschlingen 10 μl, blau, steril 2900254437
corning serologische Pipetten 50ml 14303
weighing dish 500 ST 1884.1 Carl Roth GmbH & Co.KG
filter paper Z274836-1PAK Sigma-Aldrich Chemie GmbH
inoculation loop 10µl 2900254437 NEOLAB GMBH

Table 10: Plasmides

Plasmide Supplier Catalog Number

Table 11: Buffers

NEBuffer Pack #4 (green) New England Biolabs GmbH B7004 S
NEBuffer Pack #1 (yellow) New England Biolabs GmbH B7001 S
NEBuffer Pack for T4 DNA Ligase New England Biolabs GmbH B0202 S
NEBuffer Pack #2 (blue) New England Biolabs GmbH B7002 S
NEBuffer Pack #3 (red) New England Biolabs GmbH B7003 S
TAE - Puffer (50X) für die Molekularbio VWR International GmbH A4686.1000
Gel Loading Buffer Sigma-Aldrich G2526-5ML
Tris Acetate-EDTA buffer Sigma-Aldrich T9650-1L

Table 12: other chemicals

Isopropyl B-D-Thiogalactopyranoside 1 ST Sigma-Aldrich Chemie GmbH I5502-1G
Dimethyl Sulfoxide PCR Reagent Sigma-Aldrich Chemie GmbH D9170-1VL
Glycerol Sigma Grade Sigma-Aldrich Chemie GmbH G9012-100ML
5-Bromo-4-Chloro-3-Indolyl B-D-*Galactop Sigma-Aldrich Chemie GmbH B4252-100MG
Bacteriological Agar Sigma-Aldrich Chemie GmbH A5306-250G
L-Plus-Arabinose Crystalline Sigma-Aldrich Chemie GmbH A3256-25G
Calciumchlorid Dihydrat Th. Geyer GmbH & Co KG SA/00223506/000500
Malt extract from starch digestion Sigma-Aldrich Chemie GmbH M0383-100G
D(+)-Saccharose, ACS, for Micro-Biology Sigma-Aldrich Chemie GmbH 84100-1KG
Dimethyl Sulfoxide Plant cell culture*TE D4540-100ML
Sodium Hydroxide Anhydrous Pellets Th. Geyer GmbH & Co.KG SA/S5881/000500
TRIZMA(R) Hydrochloride PH 3.5-5.0 Sigma-Aldrich Chemie GmbH T6666-50G
L-Glutamine 200 MM sterile Sigma-Aldrich Chemie GmbH G7513-20ML
Ethanol 96% Vergällt Carl Roth GmbH & Co.KG T171.3
Propanol-2 Sigma-Aldrich Chemie GmbH 59309-1L
Natriumdodecylsulfat,SDS,99%, ultra pure 13904
Gold(III)-chlorid Carl Roth GmbH & Co.KG 5624.1
Pro-Leu Sigma-Aldrich Chemie GmbH P1130-1G
Salpetersäure 65% p.a. Iso Carl Roth GmbH & Co.KG X943.1
mops, sodium Sigma-Aldrich Chemie GmbH M9024-25G
L-Glutamine Sigma-Aldrich Chemie GmbH G7513-100ML
Chelating Resin Sigma-Aldrich Chemie GmbH C7901-50G
potassium hydroxide in platellets 6751.3
hydrochloric acid 37% 4625.1
pyruvic acid Sigma-Aldrich Chemie GmbH 107360-25G
Fmoc-Orn(BOC)-OH 96.0 % 47560-5G-F
Glycerol >99,5% Sigma-Aldrich Chemie GmbH G9012-1L
Water Molecular Biology Reagent Sigma-Aldrich Chemie GmbH W4502-1L
Acetonitrile Sigma-Aldrich Chemie GmbH 34967-1L
ascorbic acid 99% Sigma-Aldrich Chemie GmbH A92902-100G

Materials used by IGEM Heidelberg 2010

  • good documented

https://2010.igem.org/Team:Heidelberg/Notebook/Material

Retrieved from "http://2013.igem.org/Team:Heidelberg/Templates/Materials_list"