Team:Heidelberg/Templates/Materials list
From 2013.igem.org
Kits
Kit | Supplier | Catalog Number |
---|---|---|
MinElute® PCR Purification Kit (250) | QIAGEN | 28006 |
Plasmid Plus Maxi Kit (25) | QIAGEN | 12963 |
Plasmid Plus Midi Kit (25) | QIAGEN | 12943 |
QIAEX II® Gel Extraction Kit (500) | QIAGEN | 20051 |
QIAGEN® Plasmid Plus Midi Kit (100) | QIAGEN | 12945 |
QIAquick® Gel Extraction Kit (250) | QIAGEN | 28706 |
QIAquick® Nucleotid Removal Kit (250) | QIAGEN | 28306 |
QIAquick® PCR Purification Kit (250) | QIAGEN | 28106 |
QIAprep® Spin Miniprep Kit (250) | QIAGEN | 27106 |
QIAprep Spin Miniprep Columns | QIAGEN | 27115 |
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 |
Enzymes
Enzyme | Supplier | Catalog Number |
---|---|---|
DreamTaq Green PCR Master Mix (2X) | Thermo Fisher Scientific Biosciences GmbH | K1081 |
Dream Taq PCR MM | Fermentas Life Sciences | K1071 |
Gibson Assembly® Master Mix | New England Biolabs | E2611 S |
Lysozyme from chicken egg white | Sigma-Aldrich Chemie GmbH | L4919-500MG |
Phusion Flash High-Fidelity PCR MAster Mix | Biozym Scientific GmbH | F-548L |
Phusion® High-Fidelity PCR Master mix | New England Biolabs | M0531 L |
T4 DNA Ligase | New England Biolabs GmbH | M0202 S |
2x PCR Master mix Solution (iTaq) | HISS DIAGNOSTICS GmbH | 25028 |
Pronase from Streptomyces griseus | Sigma-Aldrich Chemie GmbH | P6911-100MG |
Restriciton Enzymes
Enzyme | Supplier | Catalog Number |
---|---|---|
BamI | New England Biolabs | R3136 S |
BgIII | New England Biolabs | R0144 S |
DpnI | New England Biolabs | R0176 S |
EcoRI | New England BioLabs | R0101S |
EcoRI-HF | New England Biolabs | R3101 |
HindIII-HF | New England Biolabs | R3104 S |
KpnI-HF | New England Biolabs | R3142 S |
MfeI-HF | New England Biolabs | R3589 S |
NheI-HF | New England BioLabs | R3131 S |
NotI-HF | New England BioLabs | R3189 S |
PacI | New England Biolabs | R0547 S |
PstI-HF | New England Biolabs | R3140 S |
PvuI-HF | New England BioLabs | R3150S |
SalI-HF | New England Biolabs | R3138 S |
SpeI-HF | New England BioLabs | R3133 S |
XbaI | New England BioLabs | R0145 S |
Bacterial Strains
Strain | Supplier | Catalog Number |
---|---|---|
Delftia acidovorans DSM-39 | DSMZ | DSM 50251 |
Delftia acidovorans SPH-1 | DSMZ | DSM 14801 |
E. coli DH10ß | New England Biolabs | C3019 |
E. coli Top10 | invitrogen | C404010 |
Photorhabdus laumondii luminescens | DSMZ | DSM 15139 |
Streptomyces lavendulae lavendulae | DSMZ | DSM 40708 |
Streptomyces mobaraensis | DSMZ | DSM 40903 |
Antibiotics and Media Supplements
Antibiotic | Supplier | Catalog Number | Concentration stock solution | Dilution | Solvent |
---|---|---|---|---|---|
Ampicillin anhydrous crystalline | Sigma-Aldrich Chemie GmbH | A9393-5G | 100mg/ml | 1:1000 | H2O |
Ampicillin sodium crystalline | Sigma-Aldrich Chemie GmbH | A9518-5G | 100mg/ml | 1:1000 | H2O |
Chloramphenicol crystalline | Sigma-Aldrich Chemie GmbH | C0378-5G | 30mg/ml | 1:3000 | Ethanol |
Kanamycinsulfat Gemisch der Komponentena | Sigma-Aldrich Chemie GmbH | 60615-5G | 50mg/ml | 1:1000 | H2O |
Tetracycline | Sigma-Aldrich Chemie GmbH | T7660 | 10mg/ml | 1:1000 | Ethanol |
Propionic acid sodium insect cell*culture | Sigma-Aldrich Chemie GmbH | P5436-100G | 100mM | 10mM | H2O |
Bacitracin | Sigma-Aldrich Chemie GmbH | B0125-50KU | - | - |
Medium
Media | Supplier | Catalog Number |
---|---|---|
SOC Outgrowth Medium | New England Biolabs GmbH | B9020 S |
LB BROTH BASE | Th. Geyer GmbH & Co KG | SA/L3022/001000 |
LB Broth powder | Sigma-Aldrich Chemie GmbH | L3022-1KG |
M9 minimal salts | SERVA | 48505.01 |
Electrophoresis
Agarose molecular Biology Reagent | Th. Geyer GmbH & Co KG | SA/A9539/000050 | 0,5% | H2O |
Agarose for routine use | Sigma-Aldrich Chemie GmbH | A9539-100G | - | - |
Miscellaneous 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 |
Primers and Oligos
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 | TTTTTTAATTAATCACACAGGAAAGTAC TAGATGGACCGTGGCCGCCTGC |
DN13:Screen_DelH_fw | 15-05-2013 | PCR Screening for presence of DelH insert | GTAAACCCACTGGTGATACCATTC |
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 | TCGCGGCGATCCGGTACTGCGCCTCTGTT GAACATCTGATATTCTCCTCTTTAATCG ACAGATTGTGTGAAATTGTTATCCGCTCAC |
FS_02: DelAG_1_fw | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | TTCAACAGAGGCGCAGTACCGGATC |
FS_03: DelAG_1_rv | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | GTCGGAGACGATGTGGTGCATCAC |
FS_04: DelAG_2_fw | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | CTGCAGGCCAATGAGCACATCCTG |
FS_05: DelAG_2_rv | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | CACAGGTGGTAGATGGCGTC |
FS_06: DelAG_3_fwG | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | ATTGCGAGGACTTGCTCGATG |
FS_07: DelAG_3_rv | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | TTTGCTGCAGCGCCAGCACATCGAG |
FS_08: DelAG_4_fw | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | GTACGGCCTATCACATCAGCG |
FS_09: DelAG_4_rv | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | GAAGCTCAGCAGGTTGGGCGAGACG |
FS_10: DelAG_5_fw | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | GAATTTTGTTCCACCACCTGCTG |
FS_11: DelAG_5_rv | 2013-06-28 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer with overhang to DelOP | CTTGAGCAGGCGCAGTACCTCGGAGGG CGGTCGGCTGGCGTTTTCCATGATT CAGGTTTCCTGTGTGAAGCTCATCTCAGATA TCTCCCAGAGTTTCGAGAAAG |
FS_11: DelAG_5_short_rv | 2013-05-07 | Amplification of DelAG from Delftia acidovorans genome Gibson Primer | TCAGATATCTCCCAGAGTTTCGAGAAAG |
FS_12: DelOP_fw | 2013-06-28 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer | GAATCATGGAAAACGCCAGCCGAC |
FS_13: DelOP_rv | 2013-06-28 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer with overhang to DelL | CAATGTTGGAGGGGCCGAAGCCGATGCCGATC AGCGGGTGGGTTTGCATGGAAGGTC CTTTCATTGGGTCGATGCGTCCAGTGT CACACCGTGGTGTCTGCAGGCG |
FS_13: DelOP_short_rv | 2013-05-07 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer with overhang to DelL | TCACACCGTGGTGTCTGCAGGCG |
FS_14: DelL_fw | 2013-06-28 | Amplification of DelL from Delftia acidovorans genome Gibson Primer | CAAACCCACCCGCTGATCGGCATC |
FS_15: DelL_mRFP_pSB4K5_rv | 2013-06-28 | Amplification of DelL Gibson Primer with overhang to BBa_J04450 | GAAACGCATGAACTCTTTGATAACGTCT TCGGAGGAAGCCATCTAGTATTTCTCCT CTTTCTCTAGTATCAGTCCTGCAGCG CCAGCTGTTCTGTG |
FS_15: DelL_mRFP_pSB4K5__short_rv | 2013-05-07 | Amplification of DelL Gibson Primer with overhang to BBa_J04450 | TCAGTCCTGCAGCGCCAGCTGTTCTGTG |
FS_16: mRFP_pSB4K5_fw (Del) | 2013-06-28 | Amplification of pSB4K5 from iGEM Distribution Gibson Primer | GCTTCCTCCGAAGACGTTATC |
FS_20: DelF_fw | 2013-07-13 | Amplification of DelF from Delftia acidovorans genome Gibson Primer | GACTTGCTCGATGCGGTGCAG |
FS_21: DelF_fw | 2013-07-13 | Amplification of DelF from Delftia acidovorans genome Gibson Primer | GACGCCATCTACCACCTGTG |
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 |
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 | GATGAACAAATTCCGCCGCCCTGCGTCA TCTCAGATATCTCCCAGAGTTTCGAGAAAG |
FS_24: DelAE_rv | 2013-07-13 | Amplification of DelAE from Delftia acidovorans genome Gibson Primer | CAGAAGAATTCCCAGAAGGAGATGTCGAAG |
FS_25: DelEF_fw | 2013-07-13 | Amplification of DelEF from Delftia acidovorans genome Gibson Primer | ACACGGTGCTGCAGAAAACGCCCTTC |
FS_26: DelFG_rv | 2013-07-13 | Amplification of DelFG from Delftia acidovorans genome Gibson Primer | GAATTCATCCACGATGATCTGCATG |
FS_27: DelOP_rv | 2013-08-02 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer | CTTTGGGCCGTGCCGGTTTTTGAGATAC |
FS_28: DelOP_rv | 2013-08-02 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer | GTTTTTGAGATACGCGCGTTGTCAC |
FS_29: DelOP_rv | 2013-08-02 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer | TTCCCCCTCTCTTTCTCGCTTC |
FS_30: DelOP_rv | 2013-08-02 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer | CCGCTTCCCCCTCTCTTTCTCGCTTC |
FS_31: DelOP_fw | 2013-08-02 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer | GTTGGCGAGTTCAAGAAATG |
FS_32: DelOP_fw | 2013-08-02 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer | TCCTTCAGGTGTGCGGCAGACAAG |
FS_33: DelOP_fw | 2013-08-02 | Amplification of DelOP from Delftia acidovorans genome Gibson Primer | TTCTTCGTGATGACGCAGGGCGGCGGAATTTGTTC |
FS_35: DelG_fw | 2013-08-05 | Amplification of DelG from Delftia acidovorans genome Gibson Primer | CATGCAGATCATCGTGGATGAATTC |
FS_45: pSB4K5_fw | 2013-08-02 | 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 | TCAGTCCTGCAGCGCCAGCTGTTCTG TGCTTTCGTTTTATTTGATGCCT |
FS_47_screening_BB_AF_fw | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GTTGGCCGATTCATTAATGC |
FS_48_screening_BB_AF_rv | 2013-08-02 | Primer for screening/sequencing of pFSN construct | TAACGGTATCGGTATCGCTTTG |
FS_49_screening_AFI_AFII_fw | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GTTTCTCTGGAAGATGGATAC |
FS_50_screening_AFI_AFII_rv | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GTTGACGAAAAAGCCGACCAC |
FS_51_screening_AF_FG(21-26)_fw | 2013-08-02 | Primer for screening/sequencing of pFSN construct | TGGATATCGACTGGACTGCCTG |
FS_52_screening_AF_FG(21-26)_rv | 2013-08-02 | Primer for screening/sequencing of pFSN construct | TGCACCACATCGACGAAACGG |
FS_53_screening_FG(21-26)_G_fw | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GTACGGCCTATCACATCAGCG |
FS_54_screening_FG(21-26)_G_rv | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GAACCTGGGTGTTCACGAAAAAGCC |
FS_55_screening_G_OP_fw | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GTATCTCTACATGCATCGCTAC |
FS_56_screening_G_OP_fw | 2013-08-02 | Primer for screening/sequencing of pFSN construct | AGGACATTTTCCGCACCCCG |
FS_57_screening_G_OP_rv | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GCTGGCGTTTTCCATAAG |
FS_58_screening_OP_L_fw | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GAACAACTTCCAGCACAGCCTGTTC |
FS_59_screening_OP_L_rv | 2013-08-02 | Primer for screening/sequencing of pFSN construct | CGTTGAAGATTTCGTTGACG |
FS_60_screening_L_BB_fw | 2013-08-02 | Primer for screening/sequencing of pFSN construct | CATCTTCAAGGTGTTCTATGAAC |
FS_61_screening_L_BB(with_mRFP)_rv | 2013-08-02 | Primer for screening/sequencing of pFSN construct | CAGTTTAACTTTGTAGATGAAC |
FS_66: DelH_rv | 2013-08-26 | Amplification of DelH from Delftia acidovorans genome Gibson Primer | TGGGCATTCACCGCATCGATC |
FS_67: DelH_fw | 2013-08-26 | Amplification of DelH from Delftia acidovorans genome Gibson Primer | CTTCACGTTGATTGCGCATG |
FS_68: DelH_rv | 2013-08-26 | Amplification of DelH from Delftia acidovorans genome Gibson Primer | CAGAAGAACTCCCAGACCGAC |
FS_69: DelH_fw | 2013-08-26 | Amplification of DelH from Delftia acidovorans genome Gibson Primer | GACACCGTTCAGCTTCGATG |
FS_70: DelH_rv | 2013-08-26 | Amplification of DelH from Delftia acidovorans genome Gibson Primer | GAAGCTGCTCCGCTGATAGAT |
FS_71: DelH_fw | 2013-08-26 | Amplification of DelH from Delftia acidovorans genome Gibson Primer | ATGTGCTGTCGCTCAAGATG |
FS_72_SR_02_fw | 2013-08-30 | Screening of pFHFSN | ATGTGCTGTCGCTCAAGATG |
FS_73_SR_03_fw | 2013-08-30 | Screening of pFHFSN | GTGCTGTTTGGCCGTATG |
FS_74_SR_04_fw | 2013-08-30 | Screening of pFHFSN | ATCAGGTGCTGAGCTACGAC |
FS_75_SR_05_fw | 2013-08-30 | Screening of pFHFSN | CTGTTCATCAACACCTTGCC |
FS_76_SR_06_rv | 2013-08-30 | 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 | GCGATTTGGCGCAGGCGGCCACGGTC CATCTAGTATTTCTCCTCTTTC |
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 | GATTTGGCGCAGGCGGCCACGGTCCA TCTAGTACTTTCCTGTGTGACTCTAG AGCTAGCATTGTACCTAGGACTGAGCTAG CCATAAACTCTAGAAGCGGCCGCGAATTC |
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 | GCTCAGTCCTAGGTACAATGCTAGCT CTAGAGTCACACAGGAAAGTACTAGA TGGACCGTGGCCGCCTGCG |
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 | GCGATTTGGCGCAGGCGGCCACGGTCC ATCTAGTATTTCTCCTCTTTC |
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 | GGCGATTTGGCGCAGGCGGCCACGG TCCATGTACTTCGAGTCACTAAGGGCTAAC |
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 | CGCTGGAGTACGCGCTGGACTGA GATCCCAGGCATCAAATAAAACG |
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 | CTCAGTCCTAGGTACAATGCTAGCTCTAGA GTCACACAGGAAAGCAGTACACTGGCT GTGTATAAGGGAG |
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 | GTTCACCGACAAACAACAGATGA TCCGCGTGGATCCGGCTTAC |
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_rev | 2013-07-09 | Gibson-Primer DelH | TGCTGCGCCTGCATACGGCCAAACA |
HM03:DelH_Gib1.2_fw | 2013-07-09 | Gibson-Primer DelH | AGCGGCAGGGACGACGTGGT |
HM04:DelH_Gib1.2_rev | 2013-07-09 | Gibson-Primer DelH | CATAGAGGTTGTAGAGA |
HM05:DelH_Gib2.1_fw | 2013-07-09 | Gibson-Primer DelH | AGAACGCCGTCTTCAGGCTCCTG |
HM06:DelH_Gib2.1_rev | 2013-07-09 | Gibson-Primer DelH | CAATGCTTTG CCGCTCGAA |
HM07:DelH_Gib2.2_fw | 2013-07-09 | Gibson-Primer DelH | TCGCCACGGCAGCTGTTCGA |
HM08:DelH_Gib2_end_rev | 2013-07-09 | Gibson-Primer DelH | TCAGTCCAGCGCGTACTCCAG |
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 | TTGCAAAGCGCTCGGCGATTTGGCGCAGGCG GCCACGGTCCATTTAACTTTCTCCTC TTTAATACTTTGAGCTAGCCCAA AAAAACGGTATGGAGAAACAGTAGAGAGTT |
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 |
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 | TCGGCGATTTGGCGCAGGCGGCCACGGTCC ATCTAGTATTTCTCCTCTTTCTCTAGTATGTGTG |
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 | ATTGGCGCTGGAGTACGCGCTGGACTG ATCAAAGTATTAAAGAGGAGAAAGT TAAATGGCTTCCTCCGAAGACGTTATCAAAGAG |
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 |
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 |
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 |
HM16:tetR_pSB6A1_fw | 2013-08-16 | Gibson-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 | GATTTGGCGCAGGCGGCCAC GGTCCATCTAGTATTTCTCCTCTTTC |
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 | TCCGGCGCCGCCATCTGCGAGAAT CACTAGCAACTTCATTAAAGAGGAGAAATACTAG 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)_rv | 2013-08-02 | Primer for screening/sequencing of pFSN construct | GTTCACCGACAAACAACAGATAAAACG |
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_l | TTGTGTAATATTATTTTCTAACAT AAGCGACTGCTGGTTCGCG |
AR05 | 2013-09-17 | l-Ind_fw | AACGCGAACCAGCAGTCGCTT ATGTTAGAAAATAATATTACACAATGT |
AR06 | 2013-09-17 | lTycC4dC_fw | ATG CGCAACTATCCGGTCGAGACG |
AR07 | 2013-09-17 | TycC4dCom_short-BB_rev | GACGAGCGGCAGCAAGTGCTTCAT CTAGTATTTCTCCTCTTTCTCTAGTATGTG |
AR08 | 2013-09-17 | TycC4dCom_long-BB_rev | CGTCTCGACCGGATAGTTGCGCAT 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 | GCAT GAATTCGCGGCCGCTTCTAG A TGATCGCCGCCCTCCTGCCCTCC |
KH18_BBa-svp_Svert_rv | 2013-08-09 | submitting svp_Svert to registry | 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 | TGGCAGCAGAGCAGCGATCATCTAGTATTT CTCCTCTTTCCTCTAGATCATCATTCCC CCAGCAGGTATCTAAT |
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 | GATAGGACTGCCGTGCCATAATAA CGCTGATAGTGCTAGTGTAGATCGC |
NI10:pSB1C3_PstI_rv | 2013-06-04 | for linearizing, amplifying pSB1C3 | GCTTGTTTCCTGCAGTGTCATCTAG TATTTCTCCTCTTTCTCTAGTATGTG |
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 | CAGGAAACCTAAATAATGATC GCTGCTCTGCTGCCAAGTTGG |
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 |
RB06:BpsA-RBS2_rv | 2013-07-01 | Gibson assembly of pSB1C3 bpsA svp | CTTTCCTGTGTGA AAGCTT TCA TCACTCCCCGAGCAGATATCG |
RB07:RBS2-svp_fw | 2013-07-01 | Gibson assembly of pSB1C3 bpsA svp | CTTTCACACAGGAAAG TAAATA ATGGCTGCTCTTCTTCCTAGTTGGGC |
RB08:svp-pSB1C3_rv | 2013-07-01 | Gibson assembly of pSB1C3 bpsA svp | CTATCAGCGTTATTA AAGCTT TCATCA TGGCACGGCAGTCCTATCG |
RB09:pSB1C3_fw | 2013-07-01 | Gibson assembly of pSB1C3 bpsA svp | AAGCTT TAATAACGCTGATAGTGCTAGTGTAGATCGC |
RB10:pSB1C3-RBS1_rv | 2013-07-01 | Gibson assembly of pSB1C3 bpsA svp | CTAGTA TTTCTCCTCTTT CTCTAGTATGTGTG |
RB11:BpsA_Taka_fw | 2013-07-08 | colony PCR bpsA Takahashi | CAT ATGACTCTTCAGGAGACCAGCGTGCTC |
RB12:BpsA_Taka_rv | 2013-07-08 | colony PCR bpsA Takahashi | AAG CTTCTCGCCGAGCAGGTAGCGGATGTG |
RB13:svp_Taka_fw | 2013-07-08 | colony PCR svp Takahashi | CAT ATGATCGCCGCCCTCCTGCCCTCCTG |
RB14:svp_Taka_rv | 2013-07-08 | colony PCR svp Takahashi | CTCGAGCGGGACGGCGGTCCGGTCGTCCGC |
RB15:svp_Sanch_fw | 2013-07-08 | PCR svp Sanchez | TATA ATGCATGCTCGCCGCCCTCCCC |
RB16:svp_Sanch_rv | 2013-07-08 | PCR svp Sanchez | TTAAGATCTCGGGACGGCGGTCCGGTC |
RB17:sfp_fw | 2013-07-08 | sfp extraction from BAP1 | ATGAAGATTTACGGAATTTATATGG |
RB18:sfp_rv | 2013-07-08 | sfp extraction from BAP1 | TTATAAAAGCTCTTCGTACGAGACC |
RB19:entD_fw | 2013-07-08 | entD extraction from E.coli Lambalot | TAAATA ATGGTCGATATGAAAACTACGC |
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 |
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 | GGCGGATATCCTATGAGTTTGAGATTGC | |
RB81:indCdT_3_rv | 2013-09-06 | TCCATTGGCCGTCAAAGGTAATTTATCG | |
RB82:bpsA-T3_fw | 2013-09-06 | CGATAAATTACCTTTGACGGCCAATGGA AAGATCGATGTGAAAGCACTGGCCGCTTCTGACC | |
RB83:bpsA-T3_rv | 2013-09-06 | GCAATCTCAAACTCATAGGATATCCGCC CAGACCTGGCCAGCAGATCACAGG | |
RB84:indCdT_4_fw | 2013-09-06 | TTTGAAGTTGCATACCAGCTTGAACAAGC | |
RB85:bpsA-T4_rv | 2013-09-06 | GCTTGTTCAAGCTGGTATGCAACTTCAAA TGCCACGCGAGCTCCGAAGCTATATCC | |
RB86:indC-T2_fw | 2013-09-06 | TTACAGATACTTTTTCAATCTCCTAATATAGCAG | |
RB87:indC-T2_rv | 2013-09-06 | TACTTCCATCCAAATTTTTCCAAGC | |
RB88:bpsA-T2_fw | 2013-09-06 | GCTTGGAAAAATTTGGATGGAAGTA CTGAGACGCGAAAATGCTAGTGTCC | |
RB89:bpsA-T2_rv | 2013-09-06 | TTAGGAGATTGAAAAAGTATCTGTAA AGGCAGGGACACTCCCAGTCTAGCATTCAG | |
RB90:indC-T-screen_fw | 2013-09-13 | GATCAATGCGGCCTTTAATATTCG | |
RB91:bpsA-T-screen_fw | 2013-09-13 | GGAACTGAATGCTAGACTGGGAG | |
RB92:entF-T3_fw | 2013-09-16 | CGATAAATTACCTTTGACGGCCAATGGA AAGCTGGATCGCAAAGCCTTACCGTTG | |
RB93:tycA1-T3_fw | 2013-09-16 | CGATAAATTACCTTTGACGGCCAATGGA AAGATCGACCGCAAAGCGTTGC | |
RB94:tycC6-T3_fw | 2013-09-16 | CGATAAATTACCTTTGACGGCCAATGGA AAAGTGGATCGCAAAGCTTTG | |
RB95:delH4-T3_fw | 2013-09-16 | CGATAAATTACCTTTGACGGCCAATGGA AAGCTGGACCGGCAGGCCCTG | |
RB96:delH5-T3_fw | 2013-09-16 | CGATAAATTACCTTTGACGGCCAATGGA AAGCTTGACCGTGGTGCGCTG | |
RB97:plu2642-T3_fw | 2013-09-16 | CGATAAATTACCTTTGACGGCCAATGGA AAAATCGATTTCGACACATTACAAG | |
RB98:plu2670-T3_fw | 2013-09-16 | 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 | GTTCTGGCATCCTTCTTAATTAACAT TGCAGCGTGTTTCACTTGATCGATAAC | |
RB102:indC_Ox_fw | 2013-09-24 | ATGTTAATTAAGAAGGATGCCAGAAC | |
RB103:plu2642_valInd_fw | 2013-09-24 | CAGCCTGCCAGCAGCAAAACG ATGCAATCAACTCTCCCAATAATAAAATGG | |
RB104:plu2642_valInd_rv | 2013-09-24 | CAT CGTTTTGCTGCTGGCAGGCTG | |
RB105:tycC2_T3_fw | 2013-09-24 | CGATAAATTACCTTTGACGGCCAATGGA AAAGTGGATCGCAAGGCATTG | |
RB106:tycC2_T1_rv | 2013-09-24 | AGAGTCTGTCTGTTCAATCCACTT GGCCAGGCCCGCGATCGTTGG | |
RB107:tycC2_A_fw | 2013-09-24 | CATACTAGAG AAAGAGGAGAAA GGTACC ATGACGGAAGCGGAAAAACGCACACTCCTTC | |
RB108:tycC2_A_rv | 2013-09-24 | GTTCTGGCATCCTTCTTAATTAACAT GACGACCGCTTTGACCGCTTCATG | |
RB109:tycC2_valInd_fw | 2013-09-24 | CAGCCTGCCAGCAGCAAAACG ATGACGGAAGCGGAAAAACGCACACTCCTTC | |
RB110:plu2642_TE_fw | AGTGGATTGAACAGACAGACTCT AAAATCGTTGAAGGTGAAGTAACCAG |
Plasmids
Delftibactin
Name | Date | Brief Description | Genotype | Plasmid Map | GenBank-File |
---|---|---|---|---|---|
pIK1 | 2013-08-12 | Methylmalonyl-CoA pathway with permeability device and sfp | pSB3C5-lacP-BBa_B0034-BBa_I746200-BBa_B0029-pcc-acca2-BBa_B0030-sfp | pIK1 | |
pIK2 | 2013-08-12 | Methylmalonyl-CoA pathway with sfp | pSB3C5-lacP-BBa_B0034-pcc-acca2-BBa_B0030-sfp | pIK2 | |
pIK7 | 2013-08-28 | Methylmalonyl-CoA pathway with sfp | pSB3K3-lacP-BBa_B0034-pcc-acca2-BBa_B0030-sfp | pIK7 | |
pIK8 | 2013-09-12 | Methylmalonyl-CoA pathway with sfp and the permeability device behind a weak promoter | pSB3C5-lacP-BBa_B0034-pcc-acca2-BBa_B0030-sfp-BBa_B0010-BBa_B0030-BBa_I746200-BBa_B0012 | pIK8 | |
pIK9 | 2013-09-18 | Methylmalonyl-CoA pathway with sfp and the permeability device behind a weak promoter | pSB3K3-lacP-BBa_B0034-pcc-acca2-BBa_B0030-sfp-BBa_B0010-BBa_B0030-BBa_I746200-BBa_B0012 | pIK9 | |
pIK10 | 2013-09-18 | Methylmalonyl-CoA pathway with sfp and the permeability device behind a weak promoter, for submission | pSB1C3-lacP-BBa_B0034-pcc-acca2-BBa_B0030-sfp-BBa_B0010-BBa_B0030-BBa_I746200-BBa_B0012 | pIK10 | |
pIK11 | 2013-09-18 | Methylmalonyl-CoA pathway with sfp, for submission | pSB1C3-lacP-BBa_B0034-pcc-acca2-BBa_B0030-sfp | pIK11 | |
pHM01 | 2013-04-29 | DelH construct assembled by Restriction Ligation Strategy | pSB6A1-BBa_K206000(AraC)-DelH-BBa_I732019(lacZ)-B0015(stop-codon) | pHM01 | |
pHM02 | 2013-07-08 | DelH construct assembled by Gibson Assemby | pSB6A1-R0010(lacI)-B0035(RBS)-DelH-B0034(RBS)-E1010(mRFP)-B0015(stop) | pHM02 | |
pHM03 | 2013-04-29 | DelH construct assembled by Gibson Assemby for existing backbone | pSB6A1-R0010(lacI)-B0034(RBS)-DelH-B0035(RBS)-E1010(mRFP)-B0015(stop) | pHM03 | |
pHM04 | 2013-08-12 | DelH construct assembled by Gibson Assemby without mRFP | pSB6A1-R0010(lacI)-B0034(RBS)-DelH-B0015(stop) | pHM04 | |
pHM05 | 2013-08-12 | DelH construct assembled by Gibson Assemby with tetracycline resistance, without mRFP | pSB6A1-R0010(lacI)-DelH-B0015(stop)-pSB1T3(tetR) | pHM05 | |
pFSN | 2013-06-28 | DelRest, Delftibactin cluster from D. Acidovorans | pSB4K5-lacZ-BBa_B0035-DelA-DelB-DelC-DelE-DelF-DelG-DelO-DelP-DelL-BBa_B0034 | pFSN | |
pFS_02 | 2013-09-26 | DelH construct assembled by Gibson Assemby with weak Promotor, weak RBS | pSB6A1-BBa_J23114-BBa_B0032-DelH | pFS_02 | |
pFS_03 | 2013-09-26 | Helper plasmid for DelH restriction cloning, DelH, flanked by KpnI and BamHI site, without promotor, | pSB4K5-DelH | pFS_03 | |
pFS_04 | 2013-09-26 | Target plasmid for DelH restriction cloning, ccdB cassette, flanked by KpnI and BamHI site | pSB6A1-BBa_J23114-BBa_B0032-ccdBcassette | pFS_04 | |
pFS_05 | 2013-09-26 | DelH construct assembled by restriction cloning using pFS_03 and pFS-04 DelH, flanked by KpnI and BamHI site, under weak promotor, | pSB6A1-BBa_J23114-BBa_B0032-DelH | pFS_05 |
Module Shuffling
Name | Date | Brief Description | Genotype | Plasmid Map | GenBank-File |
---|---|---|---|---|---|
pIK03 | 2013-mm-dd | NRPS for Pro-Leu | pSB1C3+TycB1dComdC+TycC6 | pIK03 | |
pIK04 | 2013-mm-dd | NRPS for Phe-Orn-Leu | pSB1C3+TycAdCom+TycC5-TycC6 | pIK04 | |
pIK05 | 2013-mm-dd | NRPS for Pro-Orn-Leu | pSB1C3+TycB1dComdC+TycC5-TycC6 | pIK05 | |
pPW01 | 2013-mm-dd | NRPS for Orn-Pro-Phe-Leu | pSB1C3+TycC5+TycB1dCom-C(TycB2)+TycAdCom+TycC6 | pPW01 | |
pPW02 | 2013-mm-dd | NRPS for Orn-Pro-Phe-Leu | pSB1C3+TycC5+TycB1dCom-C(TycB2)+TycAdE+TycC6 | pPW02 | |
pPW03 | 2013-08-18 | NRPS for Phe-Indigoidine | pSB1C3+TycA+C(TycC2)+IndC | pPW03 | |
pPW04 | 2013-08-18 | NRPS for Asn-Indigoidine | pSB1C3+TycC1-C(TycC2)+IndC | pPW04 | |
pPW05 | 2013-08-18 | NRPS for Val-Indigoidine | pSB1C3+TycC4+C(TycC2)+IndC | pPW05 | |
pPW06 | 2013-09-07 | NRPS for Orn-Val-Indigoidine | pSB1C3+TycC5dC+C(TycC4)+TycC4dC+C(TycC2)+indC | pPW06 | |
pPW07 | 2013-09-07 | NRPS for Orn-Val-Val-Indigoidine | pSB1C3+TycC5dC+TycC4+C(TycC4)+TycC4dC+C(TycC2)+indC | pPW07 | |
pPW08 | 2013-09-07 | NRPS for Val-Val-Indigoidine | pSB1C3+TycC4dC+C(TycC4)+TycC4dC+C(TycC2)+indC | pPW08 | |
pPW09 | 2013-09-07 | NRPS for Pro-Leu-Indigoidine | pSB1C3+TycB1dComdC+TycC6dTE+C(TycC2)+indC | pPW09 | |
pPW10 | 2013-09-07 | NRPS for Pro-Leu-Val-Indigoidine | pSB1C3+TycB1dComdC+TycC6dTE+C(TycC4)+TycC4dC+C(TycC2)+indC | pPW10 | |
pPW11 | 2013-09-07 | NRPS for Phe-Orn-Leu-Indigoidine | pSB1C3+TycAdCom+TycC5-TycC6dTE+C(TycC2)+indC | pPW11 | |
pPW12 | 2013-09-07 | NRPS for Phe-Orn-Leu-Val-Indigoidine | pSB1C3+TycAdCom+TycC5-TycC6dTE+C(TycC4)+TycC4dC+C(TycC2)+indC | pPW12 | |
pJS01 | 2013-09-18 | Helper plasmid for the tagging of NRPs with Indigoidine | pSB1C3+ccdB+C(TycC2)+indC | pJS01 |
Linker variation
Name | Date | Brief Description | Genotype | Plasmid Map | GenBank-File |
---|---|---|---|---|---|
pAR01 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with double short linkers | pSB1C3-TycC4dCom_s+TycC2_s+indC | pAR01 | |
pAR02 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with C4 short linker | pSB1C3-TycC4dCom_s+TycC2_m+indC | pAR02 | |
pAR03 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with C2 short linker | pSB1C3-TycC4dCom_m+TycC2_s+indC | pAR03 | |
pAR04 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with medium linkers | pSB1C3-TycC4dCom+TycC2+indC | pAR04 | |
pAR05 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with C2 long linker | pSB1C3-TycC4dCom_m+TycC2_l+indC | pAR05 | |
pAR06 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with C4 long linker | pSB1C3-TycC4dCom_l+TycC2_m+indC | pAR06 | |
pAR07 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with long linkers | pSB1C3-TycC4dCom_l+TycC2_l+indC | pAR07 | |
pAR08 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with C4 short and C2 long linker | pSB1C3-TycC4dCom_s+TycC2_l+indC | pAR08 | |
pAR09 | 2013-16-09 | NRPS for Valin-Asparagine-Indigoidine-Expression with C4 long and C2 short linker | pSB1C3-TycC4dCom_l+TycC2_s+indC | pAR09 |
Dinner plasmids
Name | Date | Brief Description | Genotype | Plasmid Map | GenBank-File |
---|---|---|---|---|---|
pIK6 | 2013-08-27 | pIK6: special Ilia plasmid. This plasmid does not fullfill iGEM's RFC10 but is nevertheless special. It is for expression of tasty Ham driven by a strong lacCheese inducible promoter, a RBSalami for increased fleshiness, which can only terminated by a Champignon stop codon. The pIZa backbone with Pepper resistance and high copy Onion ori will improve the users taste stimulation. Bon Appétit! | pIZa-lacCheese P-RBSalami-ham CDS-Champ Term-Pepper res-Onion Ori | pIK6 |
Domain Shuffling and PPTases
Name | Date | Brief Description | Genotype | Plasmid Map | GenBank-File |
---|---|---|---|---|---|
pMM64 | 2013-06-01 | bpsA(fus) | pETDuet-1-T7lac-bpsA(fus) | pMM64 | |
pMM65 | 2013-06-01 | svp(fus) | pETDuet-1-T7lac-svp(fus) | pMM65 | |
pKH4 | 2013-09-01 | pRB3/pMM64 derived plasmid without sfp | pSB1C3-lacP-BBa_B0034-KpnI-indC-BBa_B0029-nonsense | pKH4 | |
pKH6 | 2013-09-05 | BBa CDS sfp only for part submission | pSB1C3-sfp | pKH6 | |
pKH7 | 2013-09-05 | BBa CDS svp only for part submission | pSB1C3-svp | pKH7 | |
pKH8 | 2013-09-05 | BBa CDS entD only for part submission | pSB1C3-entD | pKH8 | |
pKH9 | 2013-09-05 | BBa CDS delC only for part submission | pSB1C3-delC | pKH9 | |
pRB1 | 2013-07-01 | bpsA svp(pMM65) | pSB1C3-lacP-BBa_B0034-bpsA-BBa_B0029-svp(pMM65) | pXXYY | pXXYY |
pRB2 | 2013-07-08 | bpsA svp | HindIII-pSB1C3-lacP-BBa_B0034-bpsA-BBa_B0029-HindIII-svp | pXXYY | pXXYY |
pRB3 | 2013-07-15 | indC sfp | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC-BBa_B0029-BamHI-sfp | pRB03 | |
pRB4 | 2013-07-15 | indC svp | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC-BBa_B0029-BamHI-svp | pRB04 | |
pRB5 | 2013-07-15 | indC svpF | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC-BBa_B0029-BamHI-svp(pMM65) | pRB05 | |
pRB6 | 2013-07-15 | indC entD | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC-BBa_B0029-BamHI-entD | pRB06 | |
pRB7 | 2013-07-15 | bpsA(pMM64) sfp | NheI-pSB1C3-lacP-BBa_B0034-KpnI-bpsA(pMM64)-BBa_B0029-BamHI-sfp | pRB07 | |
pRB8 | 2013-07-15 | bpsA(pMM64) svpF | NheI-pSB1C3-lacP-BBa_B0034-KpnI-bpsA(pMM64)-BBa_B0029-BamHI-svp(pMM65) | pRB08 | |
pRB9 | 2013-07-15 | bpsA(pMM64) svp | NheI-pSB1C3-lacP-BBa_B0034-KpnI-bpsA(pMM64)-BBa_B0029-BamHI-svp | pRB09 | |
pRB10 | 2013-07-15 | bpsA(pMM64) entD | NheI-pSB1C3-lacP-BBa_B0034-KpnI-bpsA(pMM64)-BBa_B0029-BamHI-entD | pRB10 | |
pRB11 | 2013-07-29 | pKH1-der bpsA(pMM64)-ccdb svpF | pSB1C3-lacP-BBa_B0034-bpsA(pMM64)(ccdb)-BBa_B0029-svp(pMM65) | pXXYY | pXXYY |
pRB12 | 2013-07-29 | pKH2-der bpsA(pMM64)-ccdb svpF | pSB1C3-lacP-BBa_B0034-bpsA(pMM64)(ccdb)-BBa_B0029-svp(pMM65) | pXXYY | pXXYY |
pRB13 | 2013-07-29 | pRB3-der indC-ccdb sfp | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC(ccdb)-BBa_B0029-BamHI-sfp | pXXYY | pXXYY |
pRB14 | 2013-08-12 | pRB3-der indC-ccdB sfp | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC(ccdB)-BBa_B0029-BamHI-sfp | pXXYY | pXXYY |
pRB15 | 2013-08-12 | pSB3K3 sfp | pSB3K3-lacP-BBa_B0029-BamHI-sfp-NheI | pRB15 | |
pRB16 | 2013-08-12 | pSB3K3 svp | pSB3K3-lacP-BBa_B0029-BamHI-svp-NheI | pRB16 | |
pRB17 | 2013-08-12 | pSB3K3 entD | pSB3K3-lacP-BBa_B0029-BamHI-entD-NheI | pRB17 | |
pRB18 | 2013-08-12 | pSB3K3 delC | pSB3K3-lacP-BBa_B0029-BamHI-delC-NheI | pRB18 | |
pRB19 | 2013-08-19 | pRB14-der indC-ccdB | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC(ccdB) | pXXYY | pXXYY |
pRB20 | 2013-08-19 | pRB19-der indC-HD-ccdB | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC-HD(ccdB) | pXXYY | pXXYY |
pRB21 | 2013-08-19 | pSB1C3 indC | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC | pRB21 | |
pRB22 | 2013-08-19 | pRB21-der indC-HD | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC-HD | pRB22 | |
pRB23 | 2013-08-19 | pRB22-der indC-HD | NheI-pSB1C3-lacP-BBa_B0034-KpnI-indC*dT(ccdB) | pRB23 | pRB23 |
ID | Name (derived from) | Description |
---|---|---|
-T1 | indC | native T-Domain of indC from P. luminescens |
-T2 | bpsA | native T-Domain of bpsA from S. lavendulae |
-T3 | entF | native T-Domain of entF from E. coli |
-T4 | tycA1 | native T-domain of tycA1 from Brevibacillus parabrevis |
-T5 | tycC6 | native T-domain of tycC6 from B. parabrevis |
-T6 | delH4 | native T-domain of delH4 from Delftia acidovorans |
-T7 | delH5 | native T-domain of delH5 from D. acidovorans |
-T8 | plu2642 | native T-domain of plu2642 from Photorhabdus luminescens |
-T9 | plu2670 | native T-domain of plu2670 from P. luminescens |
-T10 | synT1 | synthetic T-Domain from indC-BLAST |
-T11 | synT2 | synthetic T-Domain from indC_T-Plum-BLAST |
-T12 | synT3 | synthetic T-Domain from indC_T-BLAST |
-T13 | synT4 | synthetic T-Domain from project_5 BLAST |
-T14 | synT5 | synthetic T-Domain random 1 (indigoidine synthetases) |
-T15 | synT6 | synthetic T-Domain random 2 (project_5) |
-TTE1 | bpsA | native TTE-Domain of bpsA from S. lavendulae |
-TTE2 | entF | native TTE-Domain of entF from E. coli |
-TTE3 | tycC6 | native TTE-domain of tycC6 from B. parabrevis |
-TTE4 | delH5 | native TTE-domain of delH5 from D. acidovorans |
Acidovorax complex medium
for 1 L
- 0.5 g yeast extract (Difco-BectonDickinson)
- 1.0 g Cas amino acids (Difco)
- 2.0 g pyruvic acid
- 2.0 g L-glutamine
- 0.3 g KH2PO4
- 0.3 g MgSO4
- 2.0 g MOPS
- 4.0 g Chelex 100-resin (Sigma)
- pH adjusted to 7.2–7.3 with 5 M KOH
Important Notes:
- fill up to 900mL before adding pyruvic acid and L-glutamine
- adjust pH
- fill up to 1L
- treat with Chelex for 1h
- remove Chelex by filtration
Ampicillin Stock Solution
What | Ampicillin 100 mg / ml |
Amount | 50 ml |
Storage | -24°C freezer |
Notes | Use in 1:1000 dilution |
efficiency test
Because we daugthed the effiency of our stock solution, we prepared a little effiency test. TB or LB media were prepared with different ampicillin solutions in order to detect at which concentration bacteria cells carrying a mRFP expressing plasmid loose it.
We can conclude that our ampicilin should still be usable, since it is at least as efficient as the ampicilin stock solution optained from a different group.
Bacitracin stock solution
What | Bacitracin 5000 U/ml |
Amount | 5 ml |
Storage | -24°C freezer |
Brevibacillus parabrevis glycerol stock
What | Brevibacillus parabrevis ATCC8185 glycerol stock |
Amount | 16 x 1.3 ml |
Storage | -80°C freezer |
Chloramphenicol stock solution
What | Chloramphenicol 30 mg / ml |
Amount | 10 ml |
Storage | -24°C freezer |
Notes | Solved in 100% ethanol. Use in 1:3000 dilution |
D. acidovorans glycerol stock
What | D. acidovorans glycerol stock |
Amount | 10 x 1 ml |
Storage | -80°C freezern |
BAP 1 glycerol stock
What | E. coli BAP1 glycerol stock |
Amount | 2 x 1.3 ml |
Storage | -80°C freezer, basement |