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Team:Freiburg/plasmids
From 2013.igem.org
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| - | <tr> <th> | + | <tr> <th colspan="3"> <center> Oligos </center> </th> </tr> |
<tr> <th> oligo name </th> <th> Sequence 5'-3' </th> <th> description </th> </tr> | <tr> <th> oligo name </th> <th> Sequence 5'-3' </th> <th> description </th> </tr> | ||
<tr> <td> oIG0000 </td> <td> TGAGCAAGAGGTAAGGG </td> <td> Sequencing Primer </td> </tr> | <tr> <td> oIG0000 </td> <td> TGAGCAAGAGGTAAGGG </td> <td> Sequencing Primer </td> </tr> | ||
| Line 139: | Line 139: | ||
<tr> <td> oIG1029 </td> <td> GCGGCCGCAATTCGAACG </td> <td> H1Promotor-fwd_BBpX334a </td> </tr> | <tr> <td> oIG1029 </td> <td> GCGGCCGCAATTCGAACG </td> <td> H1Promotor-fwd_BBpX334a </td> </tr> | ||
<tr> <td> oIG1030 </td> <td> CACAAGATATATAAAGCCAAGAAATC </td> <td> U6 Promotor-rev_BBpX334a </td> </tr> | <tr> <td> oIG1030 </td> <td> CACAAGATATATAAAGCCAAGAAATC </td> <td> U6 Promotor-rev_BBpX334a </td> </tr> | ||
| - | <tr> <td> oIG2000 </td> <td> CAGACCCCGTAGAAAAGATCAAAGGATC </td> <td> Ori1-hU6-CAGenhancer- | + | <tr> <td> oIG2000 </td> <td> CAGACCCCGTAGAAAAGATCAAAGGATC </td> <td> Ori1-hU6-CAGenhancer-hSpdCas9-fwd_pX334a </td> </tr> |
| - | <tr> <td> oIG2001 </td> <td> GAAAGCTCTGAGGCACGATAGCGTCCACATCGTAGTCGGAC </td> <td> | + | <tr> <td> oIG2001 </td> <td> GAAAGCTCTGAGGCACGATAGCGTCCACATCGTAGTCGGAC </td> <td> hSpdCas9-H840A-rev_pX334a </td> </tr> |
<tr> <td> oIG2002 </td> <td> GTCCGACTACGATGTGGACGCTATCGTGCCTCAGAGCTTTC </td> <td> H840A-linkerGSAGSAG-KRAB-fwd_pX334a </td> </tr> | <tr> <td> oIG2002 </td> <td> GTCCGACTACGATGTGGACGCTATCGTGCCTCAGAGCTTTC </td> <td> H840A-linkerGSAGSAG-KRAB-fwd_pX334a </td> </tr> | ||
<tr> <td> oIG2003 </td> <td> CTTAGCATCCATGCCGGCGGAGCCTGCTGATCCGGGGCTGTCGCCTCCCAGC </td> <td> H840A-linkerGSAGSAG-KRAB-rev_pX334a </td> </tr> | <tr> <td> oIG2003 </td> <td> CTTAGCATCCATGCCGGCGGAGCCTGCTGATCCGGGGCTGTCGCCTCCCAGC </td> <td> H840A-linkerGSAGSAG-KRAB-rev_pX334a </td> </tr> | ||
| Line 167: | Line 167: | ||
<tr> <td> oIG2026 </td> <td> AAACGGCACGCTGTTTTGACCTCCATAGAAGACACCGT </td> <td> sticky end BbsI-pKM006 target_CMVmin2-sticky end BbsI-fwd_pX334 </td> </tr> | <tr> <td> oIG2026 </td> <td> AAACGGCACGCTGTTTTGACCTCCATAGAAGACACCGT </td> <td> sticky end BbsI-pKM006 target_CMVmin2-sticky end BbsI-fwd_pX334 </td> </tr> | ||
<tr> <td> oIG2027 </td> <td> TAAAACGGTGTCTTCTATGGAGGTCAAAACAGCGTGCC </td> <td> sticky end BbsI-pKM006 target_CMVmin2-sticky end BbsI-rev_pX334 </td> </tr> | <tr> <td> oIG2027 </td> <td> TAAAACGGTGTCTTCTATGGAGGTCAAAACAGCGTGCC </td> <td> sticky end BbsI-pKM006 target_CMVmin2-sticky end BbsI-rev_pX334 </td> </tr> | ||
| - | <tr> <td> oIG2028 </td> <td> CCGGTGCCACCATGTACCCATAC </td> <td> | + | <tr> <td> oIG2028 </td> <td> CCGGTGCCACCATGTACCCATAC </td> <td> hSpdCas9-H840A-fwd_pX334a </td> </tr> |
| - | <tr> <td> oIG2029 </td> <td> CGTAATCTGGAACATCGTATGGGTACATG </td> <td> Ori1-hU6-CAGenhancer- | + | <tr> <td> oIG2029 </td> <td> CGTAATCTGGAACATCGTATGGGTACATG </td> <td> Ori1-hU6-CAGenhancer-hSpdCas9-H840A-rev_pX334a </td> </tr> |
<tr> <td> oIG2030 </td> <td> AAACCAAGGTCGGAGTACTGTCCTCCGACACTAGGT </td> <td> sticky end bsbI-UAS_Zebrafish_Locus_pos58-sticky end bsbI-fwd </td> </tr> | <tr> <td> oIG2030 </td> <td> AAACCAAGGTCGGAGTACTGTCCTCCGACACTAGGT </td> <td> sticky end bsbI-UAS_Zebrafish_Locus_pos58-sticky end bsbI-fwd </td> </tr> | ||
<tr> <td> oIG2031 </td> <td> TAAAACCTAGTGTCGGAGGACAGTACTCCGACCTTG </td> <td> sticky end bsbI-UAS_Zebrafish_Locus_pos58-sticky end bsbI-rev </td> </tr> | <tr> <td> oIG2031 </td> <td> TAAAACCTAGTGTCGGAGGACAGTACTCCGACCTTG </td> <td> sticky end bsbI-UAS_Zebrafish_Locus_pos58-sticky end bsbI-rev </td> </tr> | ||
| Line 179: | Line 179: | ||
<tr> <td> oIG2037 </td> <td> CACGCGCTAGAAAAAGTTTTGGGACC </td> <td> crRNA (reverse) </td> </tr> | <tr> <td> oIG2037 </td> <td> CACGCGCTAGAAAAAGTTTTGGGACC </td> <td> crRNA (reverse) </td> </tr> | ||
<tr> <td> oIG2038 </td> <td> TAATACGACTCACTATAGGGACCGGTGCCACCATGTACC | <tr> <td> oIG2038 </td> <td> TAATACGACTCACTATAGGGACCGGTGCCACCATGTACC | ||
| - | </td> <td> | + | </td> <td> dCas9 with T7 site and Kozak-sequence (forward) </td> </tr> |
<tr> <td> oIG2039 </td> <td> AATTGCGGCCGCTCCCCAGC </td> <td> bGH Terminator (reverse) </td> </tr> | <tr> <td> oIG2039 </td> <td> AATTGCGGCCGCTCCCCAGC </td> <td> bGH Terminator (reverse) </td> </tr> | ||
<tr> <td> oIG2040 </td> <td> GCAATAGCATCACAAATTTCACAAATAAAGC </td> <td> SV40 Terminator (reverse) </td> </tr> | <tr> <td> oIG2040 </td> <td> GCAATAGCATCACAAATTTCACAAATAAAGC </td> <td> SV40 Terminator (reverse) </td> </tr> | ||
| Line 188: | Line 188: | ||
<tr> <td> oIG3001 </td> <td> GGCAGCGCCGGCTCCGGCAGCGCCTCCGGCATGGTGAGCAAGGGCGAGGAGG </td> <td> Linker1-mCherry_forw_pMZ333 </td> </tr> | <tr> <td> oIG3001 </td> <td> GGCAGCGCCGGCTCCGGCAGCGCCTCCGGCATGGTGAGCAAGGGCGAGGAGG </td> <td> Linker1-mCherry_forw_pMZ333 </td> </tr> | ||
<tr> <td> oIG3002 </td> <td> TCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATCACTTGTACAGCTCGTCCATGCCGC </td> <td> mCherry-Bb_rev_pMZ333 </td> </tr> | <tr> <td> oIG3002 </td> <td> TCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATCACTTGTACAGCTCGTCCATGCCGC </td> <td> mCherry-Bb_rev_pMZ333 </td> </tr> | ||
| - | <tr> <td> oIG3003 </td> <td> TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGCTGACAAGAAGTACAGCATCGGCCTG </td> <td> Bb-HA- | + | <tr> <td> oIG3003 </td> <td> TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGCTGACAAGAAGTACAGCATCGGCCTG </td> <td> Bb-HA-dCas9_forw_pX334a </td> </tr> |
| - | <tr> <td> oIG3004 </td> <td> GCCGGAGGCGCTGCCGGAGCCGGCGCTGCCGTCGCCTCCCAGCTGAGACAG </td> <td> | + | <tr> <td> oIG3004 </td> <td> GCCGGAGGCGCTGCCGGAGCCGGCGCTGCCGTCGCCTCCCAGCTGAGACAG </td> <td> dCas9-Linker1_rev_pX334a </td> </tr> |
| - | <tr> <td> oIG3005 </td> <td> TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGC </td> <td> Bb-HA(-NLS- | + | <tr> <td> oIG3005 </td> <td> TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGC </td> <td> Bb-HA(-NLS-dCas9)_forw_pMZ333 </td> </tr> |
<tr> <td> oIG3006 </td> <td> TCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATCAGCTGGCCTCCACTTTTCTCTTCTTTTTGGGGCTCTTGTACAGCTCGTCCATGCCGC </td> <td> mCherry-NLS-Bb_rev_pMZ333 </td> </tr> | <tr> <td> oIG3006 </td> <td> TCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATCAGCTGGCCTCCACTTTTCTCTTCTTTTTGGGGCTCTTGTACAGCTCGTCCATGCCGC </td> <td> mCherry-NLS-Bb_rev_pMZ333 </td> </tr> | ||
<tr> <td> oIG3007 </td> <td> TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGCTATGAGAGCTGCCAACCTTTGGCCAAG </td> <td> Bb-HA-ERT2_forw_pMItom </td> </tr> | <tr> <td> oIG3007 </td> <td> TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGCTATGAGAGCTGCCAACCTTTGGCCAAG </td> <td> Bb-HA-ERT2_forw_pMItom </td> </tr> | ||
| - | <tr> <td> oIG3008 </td> <td> GGACGCTTCGACCTTGCGCTTTTTCTTCGGCGAGACTGTGGCAGGGAAACCCTCTG </td> <td> ERT2-NLS(- | + | <tr> <td> oIG3008 </td> <td> GGACGCTTCGACCTTGCGCTTTTTCTTCGGCGAGACTGTGGCAGGGAAACCCTCTG </td> <td> ERT2-NLS(-dCas9)_rev_pMItom </td> </tr> |
| - | <tr> <td> oIG3009 </td> <td> GGGGAGGCAGAGGGTTTCCCTGCCACAGTCTCGCCGAAGAAAAAGCGCAAGGTC </td> <td> ERT2-NLS(- | + | <tr> <td> oIG3009 </td> <td> GGGGAGGCAGAGGGTTTCCCTGCCACAGTCTCGCCGAAGAAAAAGCGCAAGGTC </td> <td> ERT2-NLS(-dCas9)_forw_pX334a </td> </tr> |
<tr> <td> oIG3010 </td> <td> GCTGGCCTCCACTTTTCTCTTCTTTTTGGGGCTCTTGTACAGCTCGTCCATGCCGC </td> <td> mCherry-NLS3'_rev_pMZ333 </td> </tr> | <tr> <td> oIG3010 </td> <td> GCTGGCCTCCACTTTTCTCTTCTTTTTGGGGCTCTTGTACAGCTCGTCCATGCCGC </td> <td> mCherry-NLS3'_rev_pMZ333 </td> </tr> | ||
<tr> <td> oIG3011 </td> <td> AGCCCCAAAAAGAAGAGAAAAGTGGAGGCCAGCATGAGAGCTGCCAACCTTTGGCCAAG </td> <td> NLS3'-ERT2_forw_pMItom </td> </tr> | <tr> <td> oIG3011 </td> <td> AGCCCCAAAAAGAAGAGAAAAGTGGAGGCCAGCATGAGAGCTGCCAACCTTTGGCCAAG </td> <td> NLS3'-ERT2_forw_pMItom </td> </tr> | ||
| Line 204: | Line 204: | ||
<tr> <td> oIG4000 </td> <td> GATATGGCGGGGGAACAGGGG </td> <td> mCherry-fwd_pMZ700 </td> </tr> | <tr> <td> oIG4000 </td> <td> GATATGGCGGGGGAACAGGGG </td> <td> mCherry-fwd_pMZ700 </td> </tr> | ||
<tr> <td> oIG4001 </td> <td> GCCGCTGCCGGCGCCGCTGCCGGCGCTGCCCTTGTACAGCTCGTCCATGCCGC </td> <td> mCherry-rev_pMZ700/pMZ701 </td> </tr> | <tr> <td> oIG4001 </td> <td> GCCGCTGCCGGCGCCGCTGCCGGCGCTGCCCTTGTACAGCTCGTCCATGCCGC </td> <td> mCherry-rev_pMZ700/pMZ701 </td> </tr> | ||
| - | <tr> <td> oIG4002 </td> <td> GGCAGCGCCGGCAGCGGCGCCGGCAGCGGCGACAAGAAGTACAGCATCGGCCTG </td> <td> | + | <tr> <td> oIG4002 </td> <td> GGCAGCGCCGGCAGCGGCGCCGGCAGCGGCGACAAGAAGTACAGCATCGGCCTG </td> <td> dCas9-fwd_pMZ1025 </td> </tr> |
| - | <tr> <td> oIG4003 </td> <td> GTCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATTAGTCGCCTCCCAGCTGAGACAG </td> <td> | + | <tr> <td> oIG4003 </td> <td> GTCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATTAGTCGCCTCCCAGCTGAGACAG </td> <td> dCas9-rev_pMZ1025 </td> </tr> |
<tr> <td> oIG4004 </td> <td> GTTATTGGGGCTTTCTGTTTCTTGCAAATCC </td> <td> mCherry-fwd_pMU701 </td> </tr> | <tr> <td> oIG4004 </td> <td> GTTATTGGGGCTTTCTGTTTCTTGCAAATCC </td> <td> mCherry-fwd_pMU701 </td> </tr> | ||
| - | <tr> <td> oIG4005 </td> <td> GTCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATTACACCTTTCTCTTCTTCTTGGGGCTGTC </td> <td> | + | <tr> <td> oIG4005 </td> <td> GTCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATTACACCTTTCTCTTCTTCTTGGGGCTGTC </td> <td> dCas9(with NLS)-rev_pMZ1025 </td> </tr> |
| - | <tr> <td> oIG4006 </td> <td> TCGAAGCTTGGGCTGCAGGTCGACTCTAGATTACACCTTTCTCTTCTTCTTGGGGCTGTCGCCTCCCAGCTGAGACAG </td> <td> | + | <tr> <td> oIG4006 </td> <td> TCGAAGCTTGGGCTGCAGGTCGACTCTAGATTACACCTTTCTCTTCTTCTTGGGGCTGTCGCCTCCCAGCTGAGACAG </td> <td> dCas9(with NLS)-rev_pIG4004 and pIG4005 </td> </tr> |
<tr> <td> oIG5000 </td> <td> TGCCTGAAATCACTTTTTTTCAGGTTGGACCGGTGCCACCTCGCGAATGTACCCATACGATGTTCCAGATTACG </td> <td> Backbone-Nru1-HA_for_pX334a </td> </tr> | <tr> <td> oIG5000 </td> <td> TGCCTGAAATCACTTTTTTTCAGGTTGGACCGGTGCCACCTCGCGAATGTACCCATACGATGTTCCAGATTACG </td> <td> Backbone-Nru1-HA_for_pX334a </td> </tr> | ||
<tr> <td> oIG5001 </td> <td> GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCATGGCCGACAAGCAGAAGAACGGC </td> <td> (GGGGS)3_Linker-BamH1-splitC_YFP_for_BBa_K323080 </td> </tr> | <tr> <td> oIG5001 </td> <td> GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCATGGCCGACAAGCAGAAGAACGGC </td> <td> (GGGGS)3_Linker-BamH1-splitC_YFP_for_BBa_K323080 </td> </tr> | ||
| Line 220: | Line 220: | ||
<tr> <td> oIG5011 </td> <td> CTTTAATAGTGGACTCTTGTTCCAAACTGGAAC </td> <td> forward - pX334a backbone amplification (Part2) </td> </tr> | <tr> <td> oIG5011 </td> <td> CTTTAATAGTGGACTCTTGTTCCAAACTGGAAC </td> <td> forward - pX334a backbone amplification (Part2) </td> </tr> | ||
<tr> <td> oIG5012 </td> <td> GGTGGCACCGGTCCAACCTG </td> <td> reverse - pX334a backbone amplification (Part2) </td> </tr> | <tr> <td> oIG5012 </td> <td> GGTGGCACCGGTCCAACCTG </td> <td> reverse - pX334a backbone amplification (Part2) </td> </tr> | ||
| - | <tr> <td> oIG5013 </td> <td> ACTGCCTCCGCCACCACTGCCACCGCCTCCGTCGCCTCCCAGCTGAGACAG </td> <td> (GGGGS)3_Linker- | + | <tr> <td> oIG5013 </td> <td> ACTGCCTCCGCCACCACTGCCACCGCCTCCGTCGCCTCCCAGCTGAGACAG </td> <td> (GGGGS)3_Linker-dCas9_rev_pX334a </td> </tr> |
<tr> <td> oIG5014 </td> <td> GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCATGGAGAACACTGAAAACTCAGTGGATTC </td> <td> (GGGGS)3_Linker-BamH1-Caspase3_for_pMK31 </td> </tr> | <tr> <td> oIG5014 </td> <td> GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCATGGAGAACACTGAAAACTCAGTGGATTC </td> <td> (GGGGS)3_Linker-BamH1-Caspase3_for_pMK31 </td> </tr> | ||
<tr> <td> oIG5015 </td> <td> AAGTTAGTAGCTCCGCTTCCGCTGGCCTCCACCTTTCTCTTCTTCTTGGGGCTGTCGACATGATGATGATGATGATGGTGATAAAAATAGAGTTC </td> <td> P2A(first 20bp)-NLS-Sal1-Caspase3_rev_pMK31 </td> </tr> | <tr> <td> oIG5015 </td> <td> AAGTTAGTAGCTCCGCTTCCGCTGGCCTCCACCTTTCTCTTCTTCTTGGGGCTGTCGACATGATGATGATGATGATGGTGATAAAAATAGAGTTC </td> <td> P2A(first 20bp)-NLS-Sal1-Caspase3_rev_pMK31 </td> </tr> | ||
| - | <tr> <td> oIG5016 </td> <td> CTTAGCGGCAGCTTCCTTAGCGGCAGCTTCGTCGCCTCCCAGCTGAGACAG </td> <td> (EAAAK)3_Linker- | + | <tr> <td> oIG5016 </td> <td> CTTAGCGGCAGCTTCCTTAGCGGCAGCTTCGTCGCCTCCCAGCTGAGACAG </td> <td> (EAAAK)3_Linker-dCas9_rev_pX334a </td> </tr> |
<tr> <td> oIG5017 </td> <td> GAAGCTGCCGCTAAGGAAGCTGCCGCTAAGGGATCCATGGAGAACACTGAAAACTCAGTGGATTCAAAATC </td> <td> (EAAAK)3_Linker-BamH1-Caspase3_for_pMK31 </td> </tr> | <tr> <td> oIG5017 </td> <td> GAAGCTGCCGCTAAGGAAGCTGCCGCTAAGGGATCCATGGAGAACACTGAAAACTCAGTGGATTCAAAATC </td> <td> (EAAAK)3_Linker-BamH1-Caspase3_for_pMK31 </td> </tr> | ||
<tr> <td> oIG5018 </td> <td> GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCAGGCAAGCAGCAAAGTTGTCGAAGCC </td> <td> (GGGGS)3_Linker-BamH1-Casp9_for_pKJ42 </td> </tr> | <tr> <td> oIG5018 </td> <td> GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCAGGCAAGCAGCAAAGTTGTCGAAGCC </td> <td> (GGGGS)3_Linker-BamH1-Casp9_for_pKJ42 </td> </tr> | ||
| Line 240: | Line 240: | ||
<tr> <td> oIG5032 </td> <td> ACTGCCTCCGCCACCACTGCCACCGCCTCCGTCGACTGATGTTTTAAAGAAAAGTTTTTTCCGGAGG </td> <td> (GGGGS)2_Linker_Sal1_Caspase9(long)_rev_pKJ42 </td> </tr> | <tr> <td> oIG5032 </td> <td> ACTGCCTCCGCCACCACTGCCACCGCCTCCGTCGACTGATGTTTTAAAGAAAAGTTTTTTCCGGAGG </td> <td> (GGGGS)2_Linker_Sal1_Caspase9(long)_rev_pKJ42 </td> </tr> | ||
<tr> <td> oIG5033 </td> <td> </td> <td> </td> </tr> | <tr> <td> oIG5033 </td> <td> </td> <td> </td> </tr> | ||
| - | <tr> <td> oIG5034 </td> <td> GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGACAAGAAGTACAGCATCGGCCTGG </td> <td> (GGGGS)2_Linker- | + | <tr> <td> oIG5034 </td> <td> GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGACAAGAAGTACAGCATCGGCCTGG </td> <td> (GGGGS)2_Linker-dCas9-NLS_for_pX334a </td> </tr> |
| - | <tr> <td> oIG5035 </td> <td> CTTCAGCAGGCTGAAGTTAGTAGCTCCGCTTCCAGCGTAATCTGGAACATCGTATGGGTACATGCTGGCCTCCACCTTTCTCTTCTTC </td> <td> P2A(first 33bp)-HA- | + | <tr> <td> oIG5035 </td> <td> CTTCAGCAGGCTGAAGTTAGTAGCTCCGCTTCCAGCGTAATCTGGAACATCGTATGGGTACATGCTGGCCTCCACCTTTCTCTTCTTC </td> <td> P2A(first 33bp)-HA-dCas9-NLS_rev_pX334a </td> </tr> |
<tr> <td> oIG6000 </td> <td> TCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGCAGCTAGCTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTC </td> <td> pCMVfor-psB1C3 </td> </tr> | <tr> <td> oIG6000 </td> <td> TCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGCAGCTAGCTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTC </td> <td> pCMVfor-psB1C3 </td> </tr> | ||
<tr> <td> oIG6001 </td> <td> CGAGCGGTCCGCGGAACCAGCGCTACCGGACTGCCATTGCTCCGGCTGATCTGACGGTTCACTAAACCAGCTC </td> <td> pCMVrev-eGFP </td> </tr> | <tr> <td> oIG6001 </td> <td> CGAGCGGTCCGCGGAACCAGCGCTACCGGACTGCCATTGCTCCGGCTGATCTGACGGTTCACTAAACCAGCTC </td> <td> pCMVrev-eGFP </td> </tr> | ||
| Line 276: | Line 276: | ||
<tr> <td> oig6038 </td> <td> AAACGTTCAAGTCGCACAGGCCGCTCCCGCTCCCGT </td> <td> target eCFP fw </td> </tr> | <tr> <td> oig6038 </td> <td> AAACGTTCAAGTCGCACAGGCCGCTCCCGCTCCCGT </td> <td> target eCFP fw </td> </tr> | ||
<tr> <td> oig6039 </td> <td> TAAAACGGGAGCGGGAGCGGCCTGTGCGACTTGAAC </td> <td> target eCFP rv </td> </tr> | <tr> <td> oig6039 </td> <td> TAAAACGGGAGCGGGAGCGGCCTGTGCGACTTGAAC </td> <td> target eCFP rv </td> </tr> | ||
| - | <tr> <td> oIG8002 </td> <td> GCCGGCGGAGCCTGCTGATCCGGGGCTGTCGCCTCCCAGC </td> <td> | + | <tr> <td> oIG8002 </td> <td> GCCGGCGGAGCCTGCTGATCCGGGGCTGTCGCCTCCCAGC </td> <td> dCas9_GSAGSAG_rev </td> </tr> |
<tr> <td> oIG8005 </td> <td> GGACGCTTCGACCTTGCGCTTTTTC </td> <td> NLS1_pX334_rev </td> </tr> | <tr> <td> oIG8005 </td> <td> GGACGCTTCGACCTTGCGCTTTTTC </td> <td> NLS1_pX334_rev </td> </tr> | ||
| - | <tr> <td> oIG8006 </td> <td> GGATCAGCAGGCTCCGCCGGCGACAAGAAGTACAGCATCGGCCTG </td> <td> | + | <tr> <td> oIG8006 </td> <td> GGATCAGCAGGCTCCGCCGGCGACAAGAAGTACAGCATCGGCCTG </td> <td> dCas9_GSAGSAG_fw </td> </tr> |
<tr> <td> oIG8007 </td> <td> GGTGCTGGTATGACAAG </td> <td> G9a_seq_fw_pGEX2T </td> </tr> | <tr> <td> oIG8007 </td> <td> GGTGCTGGTATGACAAG </td> <td> G9a_seq_fw_pGEX2T </td> </tr> | ||
<tr> <td> oIG8008 </td> <td> GATGTTGCCATAGTAACGG </td> <td> G9a_seq_rev_pGEX2T </td> </tr> | <tr> <td> oIG8008 </td> <td> GATGTTGCCATAGTAACGG </td> <td> G9a_seq_rev_pGEX2T </td> </tr> | ||
| Line 297: | Line 297: | ||
<tr> <td> oIG8023 </td> <td> GCTGGTAGTGCTGGTTCCGCGTACAGCCGCGAGAATGTACCCATCCCCTGTGTC </td> <td> phyB(overhang)_g9a for </td> </tr> | <tr> <td> oIG8023 </td> <td> GCTGGTAGTGCTGGTTCCGCGTACAGCCGCGAGAATGTACCCATCCCCTGTGTC </td> <td> phyB(overhang)_g9a for </td> </tr> | ||
<tr> <td> oIG8024 </td> <td> CTCTAGATCACACCTTCCGCTTTTTCTTGGGGGTGTTGATGGGGGGCAGGGA </td> <td> phyB(overhang)_g9a rev </td> </tr> | <tr> <td> oIG8024 </td> <td> CTCTAGATCACACCTTCCGCTTTTTCTTGGGGGTGTTGATGGGGGGCAGGGA </td> <td> phyB(overhang)_g9a rev </td> </tr> | ||
| - | <tr> <td> oIG9100 </td> <td> TCATATCCGCGGCATGTACCCATACGATGTTCCAGATTACG </td> <td> | + | <tr> <td> oIG9100 </td> <td> TCATATCCGCGGCATGTACCCATACGATGTTCCAGATTACG </td> <td> dCas9 fw incl. HA tag and NLS </td> </tr> |
| - | <tr> <td> oIG9191 </td> <td> GATATTGGTACCCATGCCTGCTATTCTCTTCCCAATCCTC </td> <td> | + | <tr> <td> oIG9191 </td> <td> GATATTGGTACCCATGCCTGCTATTCTCTTCCCAATCCTC </td> <td> dCas9 rv incl. Bgh terminator </td> </tr> |
<tr> <td> oIG4100F </td> <td> GCTAGCTAGTTATTAATAGTAATCAATTACGGG </td> <td> wird nachgetragen </td> </tr> | <tr> <td> oIG4100F </td> <td> GCTAGCTAGTTATTAATAGTAATCAATTACGGG </td> <td> wird nachgetragen </td> </tr> | ||
<tr> <td> oIG4100R </td> <td> GAATTCGATCTGACGGTTCACTAAAC </td> <td> </td> </tr> | <tr> <td> oIG4100R </td> <td> GAATTCGATCTGACGGTTCACTAAAC </td> <td> </td> </tr> | ||
| Line 327: | Line 327: | ||
<tr> <td> crRNA tetO 1 R </td> <td> AAACACTCGACTTTCACTTTTCTCTATCACTGATGT </td> <td> </td> </tr> | <tr> <td> crRNA tetO 1 R </td> <td> AAACACTCGACTTTCACTTTTCTCTATCACTGATGT </td> <td> </td> </tr> | ||
<tr> <td> crRNA tetO 2 F </td> <td> TAAAACCTCCCTATCAGTGATAGAGAAAAGTGAAAG </td> <td> </td> </tr> | <tr> <td> crRNA tetO 2 F </td> <td> TAAAACCTCCCTATCAGTGATAGAGAAAAGTGAAAG </td> <td> </td> </tr> | ||
| - | <tr> <td> | + | <tr> <td> oIG9000 </td> <td> AGGAGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGGCTC </td> <td> ELISA CRISPR/Cas9 for EMX1 sequence, biotinylated </td> </tr> |
| + | <tr> <td> oIG9001 </td> <td> GAGCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCCTCCT </td> <td> ELISA CRISPR/Cas9 for EMX1 sequence </td> </tr> | ||
| + | <tr> <td> oIG9002 </td> <td> CAGAAGCTGGAGGAGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGGCTC </td> <td> ELISA CRISPR/Cas9 for EMX1 sequence, biotinylated </td> </tr> | ||
| + | <tr> <td> oIG9003 </td> <td> GAGCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCCTCCTCCAGCTTCTG </td> <td> ELISA CRISPR/Cas9 for EMX1 sequence </td> </tr> | ||
| + | <tr> <td> oIG9004 </td> <td> CAGAAGCTGGAGGAGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGGCTCCCATCACATC </td> <td> ELISA CRISPR/Cas9 for EMX1 sequence, biotinylated </td> </tr> | ||
| + | <tr> <td> oIG9005 </td> <td> GATGTGATGGGAGCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCCTCCTCCAGCTTCTG </td> <td> ELISA CRISPR/Cas9 for EMX1 sequence </td> </tr> | ||
| + | <tr> <td> oIG9006 </td> <td> GGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGTTTTAGAGCTATGCTGTTTTGAATGGTC </td> <td> forward primer for the insertion of the EMX1 </td> </tr> | ||
| + | <tr> <td> oIG9007 </td> <td> TTCTTCTTCTGCTCGGACTCAGGCCCTTCCGTTTTGGGACCATTCAAAACAGCATAGC </td> <td> reverse primer for the insertion of the EMX1 </td> </tr> | ||
| + | <tr> <td> oIG9008 </td> <td> GGGCTTGTACTCGGTCATGGTAAGCTTCAGCTGCTCGAGATCTAGATGGATGCAGGTCG </td> <td> reverse primer for the insertion of the EMX1 </td> </tr> | ||
| + | <tr> <td> oIG9009 </td> <td> CGACCTGCATCCATCTAGATCTCGAGCAGCTGAAGCTTACC </td> <td> forward primer for the insertion of the EMX1 </td> </tr> | ||
| + | <tr> <td> oIG9010 </td> <td> CAG CAT CGG CCT GGA CAT CGG C </td> <td> Site directed mutagenesis pcr reverse primer to reestablish the endonuclease activity by activating one inactivated nicase </td> </tr> | ||
| + | <tr> <td> oIG9011 </td> <td> GCC GAT GTC CAG GCC GAT GCT G </td> <td> Site directed mutagenesis pcr reverse primer to reestablish the endonuclease activity by activating one inactivated nicase </td> </tr> | ||
| + | <tr> <td> oIG9012 </td> <td> CAGAAGCTGGATTAAAAGTCGGCTGGTAGCGGGGAGGATCGGGGCTCCCATCACATC </td> <td> human VEGFA locus 1_VZ-8_fwd_pKM600, biotinylated </td> </tr> | ||
| + | <tr> <td> oIG9013 </td> <td> GATGTGATGGGAGCCCCGATCCTCCCCGCTACCAGCCGACTTTTAATCCAGCTTCTG </td> <td> human VEGFA locus 1_VZ-8_fwd_pKM600 </td> </tr> | ||
| + | <tr> <td> oIG9015 </td> <td> CAGAAGCTGGAGACCTGCTTTTGGGGGTGACCGCCGGAGCGGGGCTCCCATCACATC </td> <td> human VEGFA locus 3_VZ+434b_fwd_pKM600, biotinylated </td> </tr> | ||
| + | <tr> <td> oIG9016 </td> <td> GATGTGATGGGAGCCCCGCTCCGGCGGTCACCCCCAAAAGCAGGTCTCCAGCTTCTG </td> <td> human VEGFA locus 3_VZ+434b_fwd_pKM600 </td> </tr> | ||
| + | <tr> <td> oIG9018 </td> <td> CAGAAGCTGGACACGACTTCTTCAAGTCCGCCATGCCCGAGGGCTCCCATCACATC </td> <td> +750 Target, Forward, biotinylated </td> </tr> | ||
| + | <tr> <td> oIG9019 </td> <td> GATGTGATGGGAGCCCTCGGGCATGGCGGACTTGAAGAAGTCGTGTCCAGCTTCTG </td> <td> +750 Target, Forward </td> </tr> | ||
| + | <tr> <td> oIG9024 </td> <td> CAGAAGCTGGAAGAGCTCGTTTAGTGAACCGTCAGATCGCCGGGCTCCCATCACATC </td> <td> CMVmin 1 Target, biotinylated </td> </tr> | ||
| + | <tr> <td> oIG9025 </td> <td> GATGTGATGGGAGCCCGGCGATCTGACGGTTCACTAAACGAGCTCTTCCAGCTTCTG </td> <td> CMVmin 1 Target </td> </tr> | ||
| + | <tr> <td> oIG9041 </td> <td> CAGAAGCTGGAGTGTGCAGACGGCAGTCACTAGGGGGCGCTGGGCTCCCATCACATC </td> <td> human VEGFA locus2, biotinylated </td> </tr> | ||
| + | <tr> <td> oIG9042 </td> <td> GATGTGATGGGAGCCCAGCGCCCCCTAGTGACTGCCGTCTGCACACTCCAGCTTCTG </td> <td> human VEGFA locus 2 </td> </tr> | ||
| + | <tr> <td> oIG9043 </td> <td> CAGAAGCTGGACGTCTATATCACCGCCGACAAGCAGAAGAAGGGCTCCCATCACATC </td> <td> target_nt950 pIG9003 fwd, biotinylated </td> </tr> | ||
| + | <tr> <td> oIG9044 </td> <td> GATGTGATGGGAGCCCTTCTTCTGCTTGTCGGCGGTGATATAGACGTCCAGCTTCTG </td> <td> target_nt950 pIG9003rev </td> </tr> | ||
| + | |||
</table> | </table> | ||
<p></p> | <p></p> | ||
<table> | <table> | ||
| - | <tr> <th> | + | <tr> <th colspan="5"> <center> Plasmids </center> </th> </tr> |
<tr> <th> plasmid name </th> <th> Description </th> <th> Backbone </th> <th> Resistance </th> <th> Reference </th> </tr> | <tr> <th> plasmid name </th> <th> Description </th> <th> Backbone </th> <th> Resistance </th> <th> Reference </th> </tr> | ||
| - | <tr> <td> pIG0001 </td> <td> | + | <tr> <td> pIG0001 </td> <td> dCas9, RFC 25 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> pX33a und pIG2004 </td> </tr> |
<tr> <td> pIG0002 </td> <td> pSV40, RFC 25 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | <tr> <td> pIG0002 </td> <td> pSV40, RFC 25 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | ||
<tr> <td> pIG0003 </td> <td> SV40, RFC 25 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | <tr> <td> pIG0003 </td> <td> SV40, RFC 25 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | ||
| Line 354: | Line 378: | ||
<tr> <td> pIG0019 </td> <td> CMV </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> CMV was used from parts regestry (BBa_K747096, RFC10) </td> </tr> | <tr> <td> pIG0019 </td> <td> CMV </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> CMV was used from parts regestry (BBa_K747096, RFC10) </td> </tr> | ||
<tr> <td> pIG0020 </td> <td> BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> BGH was amplified from pGR2 with oIG0049 + oIG0050 and ligated into pSB1C3 via Gibson assembly. </td> </tr> | <tr> <td> pIG0020 </td> <td> BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> BGH was amplified from pGR2 with oIG0049 + oIG0050 and ligated into pSB1C3 via Gibson assembly. </td> </tr> | ||
| - | <tr> <td> pIG0021 a </td> <td> NLS- | + | <tr> <td> pIG0021 a </td> <td> NLS-dCas9 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0018 and pIG0001 (RFC25) </td> </tr> |
<tr> <td> pIG0022 </td> <td> CMV-HA </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0017 and pIG0019 (RFC25) </td> </tr> | <tr> <td> pIG0022 </td> <td> CMV-HA </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0017 and pIG0019 (RFC25) </td> </tr> | ||
<tr> <td> pIG0023 </td> <td> SV40-HA </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0003 and pIG0017 (RFC25) </td> </tr> | <tr> <td> pIG0023 </td> <td> SV40-HA </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0003 and pIG0017 (RFC25) </td> </tr> | ||
<tr> <td> pIG0024 </td> <td> NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0018 and pIG0020 (RFC25) </td> </tr> | <tr> <td> pIG0024 </td> <td> NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0018 and pIG0020 (RFC25) </td> </tr> | ||
| - | <tr> <td> pIG0025 </td> <td> CMV-HA-NLS- | + | <tr> <td> pIG0025 </td> <td> CMV-HA-NLS-dCas9 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0022 and pIG0021 (RFC25) </td> </tr> |
| - | <tr> <td> pIG0026 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0026 </td> <td> SV40-HA-NLS-dCas9 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0023 and pIG0021 (RFC25) </td> </tr> |
| - | <tr> <td> pIG0027 </td> <td> CMV-HA-NLS- | + | <tr> <td> pIG0027 </td> <td> CMV-HA-NLS-dCas9-Linker (3AS) </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0025 and pIG0029 (RFC25) </td> </tr> |
| - | <tr> <td> pIG0028 </td> <td> CMV-HA-NLS- | + | <tr> <td> pIG0028 </td> <td> CMV-HA-NLS-dCas9-Linker (7AS) </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0025 and pIG0030 (RFC25) </td> </tr> |
| - | <tr> <td> pIG0029 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0029 </td> <td> SV40-HA-NLS-dCas9-Linker(3AS) </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0026 and pIG0029 (RFC25) </td> </tr> |
| - | <tr> <td> pIG0030 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0030 </td> <td> SV40-HA-NLS-dCas9-Linker(7AS) </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of pIG0026 and pIG0030 (RFC25) </td> </tr> |
<tr> <td> pIG0031 </td> <td> Linker 3 AS </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of annealed oIG0045+46 and pSB1C3 RFC25 </td> </tr> | <tr> <td> pIG0031 </td> <td> Linker 3 AS </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of annealed oIG0045+46 and pSB1C3 RFC25 </td> </tr> | ||
<tr> <td> pIG0032 </td> <td> Linker (7 AS) </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of annealed oIG0051+52 and pSB1C3 RFC25 </td> </tr> | <tr> <td> pIG0032 </td> <td> Linker (7 AS) </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> Ligation of annealed oIG0051+52 and pSB1C3 RFC25 </td> </tr> | ||
| - | <tr> <td> pIG0033 </td> <td> HA-NLS- | + | <tr> <td> pIG0033 </td> <td> HA-NLS-dCas9-Linker (3AS) </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
<tr> <td> pIG0034 </td> <td> SV40 im 10er Standard </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | <tr> <td> pIG0034 </td> <td> SV40 im 10er Standard </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | ||
<tr> <td> pIG0035 </td> <td> BGH im 10er Standard </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | <tr> <td> pIG0035 </td> <td> BGH im 10er Standard </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | ||
| - | <tr> <td> pIG0036 </td> <td> HA-NLS- | + | <tr> <td> pIG0036 </td> <td> HA-NLS-dCas9-Linker (3AS)-KRAB </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0037 </td> <td> HA-NLS- | + | <tr> <td> pIG0037 </td> <td> HA-NLS-dCas9-Linker (3AS)-G9a </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0038 </td> <td> HA-NLS- | + | <tr> <td> pIG0038 </td> <td> HA-NLS-dCas9-Linker (3AS)-VP16 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0039 </td> <td> HA-NLS- | + | <tr> <td> pIG0039 </td> <td> HA-NLS-dCas9-Linker (3AS)-KRAB-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0040 </td> <td> HA-NLS- | + | <tr> <td> pIG0040 </td> <td> HA-NLS-dCas9-Linker (3AS)-G9a-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0041 </td> <td> HA-NLS- | + | <tr> <td> pIG0041 </td> <td> HA-NLS-dCas9-Linker (3AS)-VP16-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0042 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0042 </td> <td> SV40-HA-NLS-dCas9-Linker (3AS)-KRAB-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0043 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0043 </td> <td> SV40-HA-NLS-dCas9-Linker (3AS)-G9a-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0044 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0044 </td> <td> SV40-HA-NLS-dCas9-Linker (3AS)-VP16-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0045 </td> <td> CMV-HA-NLS- | + | <tr> <td> pIG0045 </td> <td> CMV-HA-NLS-dCas9-Linker (3AS)-KRAB-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0046 </td> <td> CMV-HA-NLS- | + | <tr> <td> pIG0046 </td> <td> CMV-HA-NLS-dCas9-Linker (3AS)-G9a-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0047 </td> <td> CMV-HA-NLS- | + | <tr> <td> pIG0047 </td> <td> CMV-HA-NLS-dCas9-Linker (3AS)-VP16-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
<tr> <td> pIG0048 </td> <td> pSV40-PhyB </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | <tr> <td> pIG0048 </td> <td> pSV40-PhyB </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | ||
<tr> <td> pIG0049 </td> <td> pSV40-COP1 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | <tr> <td> pIG0049 </td> <td> pSV40-COP1 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | ||
| Line 395: | Line 419: | ||
<tr> <td> pIG0060 </td> <td> CMV-COP1-L3-VP16-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | <tr> <td> pIG0060 </td> <td> CMV-COP1-L3-VP16-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | ||
<tr> <td> pIG0061 </td> <td> CMV-COP1-L3-KRAB-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | <tr> <td> pIG0061 </td> <td> CMV-COP1-L3-KRAB-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> | ||
| - | <tr> <td> pIG0062 </td> <td> CMV-HA-NLS- | + | <tr> <td> pIG0062 </td> <td> CMV-HA-NLS-dCas9-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0063 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0063 </td> <td> SV40-HA-NLS-dCas9-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0064 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0064 </td> <td> SV40-HA-NLS-dCas9-Linker 3AS-Uvr8 </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0065 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0065 </td> <td> SV40-HA-NLS-dCas9-Linker 3AS-Piff </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0066 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0066 </td> <td> SV40-HA-NLS-dCas9-Linker 3AS-Uvr8-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
| - | <tr> <td> pIG0067 </td> <td> SV40-HA-NLS- | + | <tr> <td> pIG0067 </td> <td> SV40-HA-NLS-dCas9-Linker 3AS-Piff-NLS-BGH </td> <td> pSB1C3 </td> <td> Chloramph. </td> <td> </td> </tr> |
<tr> <td> pIG1000 </td> <td> Siehe Liste unter Ornder "Aktivierung" </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG1000 </td> <td> Siehe Liste unter Ornder "Aktivierung" </td> <td> </td> <td> </td> <td> </td> </tr> | ||
<tr> <td> pIG1001 </td> <td> Siehe Liste unter Ornder "Aktivierung" </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG1001 </td> <td> Siehe Liste unter Ornder "Aktivierung" </td> <td> </td> <td> </td> <td> </td> </tr> | ||
<tr> <td> pIG1002 </td> <td> Siehe Liste unter Ornder "Aktivierung" </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG1002 </td> <td> Siehe Liste unter Ornder "Aktivierung" </td> <td> </td> <td> </td> <td> </td> </tr> | ||
<tr> <td> pIG1003 </td> <td> Siehe Liste unter Ornder "Aktivierung" </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG1003 </td> <td> Siehe Liste unter Ornder "Aktivierung" </td> <td> </td> <td> </td> <td> </td> </tr> | ||
| - | <tr> <td> pIG2000 </td> <td> hU6 - precrRNA-site - CAG - | + | <tr> <td> pIG2000 </td> <td> hU6 - precrRNA-site - CAG - dCas9, D10A </td> <td> pX334a </td> <td> Amp, Puro </td> <td> Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong et al (Science. 2013 Jan 3. PubMed) </td> </tr> |
<tr> <td> pIG2001 </td> <td> UVR8 - KRAB - NLS </td> <td> pKM102 </td> <td> Amp, Tet </td> <td> </td> </tr> | <tr> <td> pIG2001 </td> <td> UVR8 - KRAB - NLS </td> <td> pKM102 </td> <td> Amp, Tet </td> <td> </td> </tr> | ||
<tr> <td> pIG2002 </td> <td> tetO13 - CMVmin - SEAP </td> <td> pKM006 </td> <td> Amp, ColE1 </td> <td> Reporter Plasmid, in combination with pSAM200: constitutively high expression of SEAP </td> </tr> | <tr> <td> pIG2002 </td> <td> tetO13 - CMVmin - SEAP </td> <td> pKM006 </td> <td> Amp, ColE1 </td> <td> Reporter Plasmid, in combination with pSAM200: constitutively high expression of SEAP </td> </tr> | ||
<tr> <td> pIG2003 </td> <td> tetR - VP16 </td> <td> pSAM200 </td> <td> Amp, ColE1 </td> <td> Cis-activating fusion-protein of the used tetR-System </td> </tr> | <tr> <td> pIG2003 </td> <td> tetR - VP16 </td> <td> pSAM200 </td> <td> Amp, ColE1 </td> <td> Cis-activating fusion-protein of the used tetR-System </td> </tr> | ||
| - | <tr> <td> pIG2004 </td> <td> hU6 - precrRNA-site - CAG - | + | <tr> <td> pIG2004 </td> <td> hU6 - precrRNA-site - CAG - dCas9, D10A, H840A - KRAB </td> <td> px334a </td> <td> Amp, Puro </td> <td> Zhang-plasmid, containing a H840A mutation, induced by side-directed mutagenesis (oIG2000 & 2001 / 2002 & 2003). Furtherly containing a KRAB domain, which originated from PCR amplification and subsequent Gibson Cloning of a pKM102 part (oIG2004 & oIG2005) with pX334a. </td> </tr> |
| - | <tr> <td> pIG2005 </td> <td> hU6 - precrRNA-site - CAG - | + | <tr> <td> pIG2005 </td> <td> hU6 - precrRNA-site - CAG - dCas9, D10A, H840A </td> <td> pX334a </td> <td> Amp, Puro </td> <td> Zhang-plasmid, containing a H840A mutation, induced by side-directed mutagenesis (oIG2000-oIG2001, oIG2002-oIG2007). Gibson cloned of three PCR-fragments: oIG2000-oIG2001, oIG2002-oIG2007, oIG2008-oIG2009 </td> </tr> |
| - | <tr> <td> pIG2006 </td> <td> hU6 - CMVmin-crRNA - HA - NLS - | + | <tr> <td> pIG2006 </td> <td> hU6 - CMVmin-crRNA - HA - NLS - dCas9 - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2007 </td> <td> hU6 - CMV-crRNA - HA - NLS - | + | <tr> <td> pIG2007 </td> <td> hU6 - CMV-crRNA - HA - NLS - dCas9 - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2008 </td> <td> hU6 - +230-crRNA - HA - NLS - | + | <tr> <td> pIG2008 </td> <td> hU6 - +230-crRNA - HA - NLS - dCas9 - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2009 </td> <td> hU6 - EMX-crRNA - HA - NLS - | + | <tr> <td> pIG2009 </td> <td> hU6 - EMX-crRNA - HA - NLS - dCas9 - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2010 </td> <td> hU6 - CMVmin-crRNA - HA - NLS - | + | <tr> <td> pIG2010 </td> <td> hU6 - CMVmin-crRNA - HA - NLS - dCas9 - KRAB - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2011 </td> <td> hU6 - CMV-crRNA - HA - NLS - | + | <tr> <td> pIG2011 </td> <td> hU6 - CMV-crRNA - HA - NLS - dCas9 - KRAB - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2012 </td> <td> hU6 - +230-crRNA - HA - NLS - | + | <tr> <td> pIG2012 </td> <td> hU6 - +230-crRNA - HA - NLS - dCas9 - KRAB - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2013 </td> <td> hU6 - EMX-crRNA - HA - NLS - | + | <tr> <td> pIG2013 </td> <td> hU6 - EMX-crRNA - HA - NLS - dCas9 - KRAB - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2014 </td> <td> hU6 - CMVmin-crRNA - HA - NLS - | + | <tr> <td> pIG2014 </td> <td> hU6 - CMVmin-crRNA - HA - NLS - dCas9_H840A - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2015 </td> <td> hU6 - CMV-crRNA - HA - NLS - | + | <tr> <td> pIG2015 </td> <td> hU6 - CMV-crRNA - HA - NLS - dCas9_H840A - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2016 </td> <td> hU6 - +230-crRNA - HA - NLS - | + | <tr> <td> pIG2016 </td> <td> hU6 - +230-crRNA - HA - NLS - dCas9_H840A - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2017 </td> <td> hU6 - EMX-crRNA - HA - NLS - | + | <tr> <td> pIG2017 </td> <td> hU6 - EMX-crRNA - HA - NLS - dCas9_H840A - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2018 </td> <td> hU6 - +30-crRNA - HA - NLS - | + | <tr> <td> pIG2018 </td> <td> hU6 - +30-crRNA - HA - NLS - dCas9 - NLS </td> <td> pX334 </td> <td> Amp </td> <td> - </td> </tr> |
| - | <tr> <td> pIG2019 </td> <td> hU6 - +30-crRNA - HA - NLS - | + | <tr> <td> pIG2019 </td> <td> hU6 - +30-crRNA - HA - NLS - dCas9 - KRAB - NLS </td> <td> pX334 </td> <td> Amp </td> <td> </td> </tr> |
| - | <tr> <td> pIG2020 </td> <td> hU6 - +30-crRNA - HA - NLS - | + | <tr> <td> pIG2020 </td> <td> hU6 - +30-crRNA - HA - NLS - dCas9_H840A - NLS </td> <td> pX334 </td> <td> Amp </td> <td> </td> </tr> |
| - | <tr> <td> pIG2021 </td> <td> hU6 - Vegf II-crRNA - HA - NLS - | + | <tr> <td> pIG2021 </td> <td> hU6 - Vegf II-crRNA - HA - NLS - dCas9 - KRAB - NLS </td> <td> pX334 </td> <td> Amp </td> <td> </td> </tr> |
| - | <tr> <td> pIG2022 </td> <td> hU6 - Vegf III-crRNA - HA - NLS - | + | <tr> <td> pIG2022 </td> <td> hU6 - Vegf III-crRNA - HA - NLS - dCas9 - KRAB - NLS </td> <td> pX334 </td> <td> Amp </td> <td> </td> </tr> |
| - | <tr> <td> pIG3001 </td> <td> | + | <tr> <td> pIG3001 </td> <td> dCas9-mCherry </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3002 </td> <td> HA-NLS- | + | <tr> <td> pIG3002 </td> <td> HA-NLS-dCas9-mCherry-NLS </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; HA-NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, the C-terminal NLS (different codon usage) was intoduced with a primer overlap; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3003 </td> <td> HA- | + | <tr> <td> pIG3003 </td> <td> HA-dCas9-mCherry-NLS </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, the C-terminal NLS (different codon usage) and HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3004 </td> <td> HA-NLS- | + | <tr> <td> pIG3004 </td> <td> HA-NLS-dCas9-mCherry </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; HA-NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3005 </td> <td> HA-ERT2-NLS- | + | <tr> <td> pIG3005 </td> <td> HA-ERT2-NLS-dCas9-mCherry-NLS-ERT2 </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the C-terminal NLS (different codon usage) and the HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3006 </td> <td> HA-NLS- | + | <tr> <td> pIG3006 </td> <td> HA-NLS-dCas9-mCherry-NLS-ERT2 </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; HA-NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the C-terminal NLS (different codon usage) was intoduced with a primer overlap; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3007 </td> <td> HA-ERT2-NLS- | + | <tr> <td> pIG3007 </td> <td> HA-ERT2-NLS-dCas9-mCherry-NLS </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the C-terminal NLS (different codon usage) and the HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3008 </td> <td> HA- | + | <tr> <td> pIG3008 </td> <td> HA-dCas9-mCherry-NLS-ERT2 </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the C-terminal NLS (different codon usage) and the HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3009 </td> <td> HA-ERT2-NLS- | + | <tr> <td> pIG3009 </td> <td> HA-ERT2-NLS-dCas9-mCherry </td> <td> pMZ333 </td> <td> Amp </td> <td> Backbone was cut out of pMZ333 by NotI and XbaI; NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR </td> </tr> |
| - | <tr> <td> pIG3010 </td> <td> - </td> <td> pX334a </td> <td> amp </td> <td> pX334a was digested with PstI to get rid of | + | <tr> <td> pIG3010 </td> <td> - </td> <td> pX334a </td> <td> amp </td> <td> pX334a was digested with PstI to get rid of dCas9 and religated </td> </tr> |
<tr> <td> pIG3011 </td> <td> SEAP-Targetsequence I </td> <td> pX334a </td> <td> amp </td> <td> pIG3010 was opened with BbsI and SEAP-Targetsequence I was inserted (oIG3013 + 14) </td> </tr> | <tr> <td> pIG3011 </td> <td> SEAP-Targetsequence I </td> <td> pX334a </td> <td> amp </td> <td> pIG3010 was opened with BbsI and SEAP-Targetsequence I was inserted (oIG3013 + 14) </td> </tr> | ||
<tr> <td> pIG3012 </td> <td> SEAP-Targetsequence II </td> <td> pX334a </td> <td> amp </td> <td> pIG3010 was opened with BbsI and SEAP-Targetsequence II was inserted (oIG3015 + 16) </td> </tr> | <tr> <td> pIG3012 </td> <td> SEAP-Targetsequence II </td> <td> pX334a </td> <td> amp </td> <td> pIG3010 was opened with BbsI and SEAP-Targetsequence II was inserted (oIG3015 + 16) </td> </tr> | ||
| Line 443: | Line 467: | ||
<tr> <td> pIG4000 </td> <td> oMZ700PhyB (1-605) - mCherry-sv40pA </td> <td> </td> <td> bla,ColE1 </td> <td> mCherry was first PCR amplified from pMZ300 with oligos oMZ701/oMZ706, further amplified with oligos oMZ703/oMZ707, and Gibson-cloned into pMZ331 cut with XbaI/EcoRI </td> </tr> | <tr> <td> pIG4000 </td> <td> oMZ700PhyB (1-605) - mCherry-sv40pA </td> <td> </td> <td> bla,ColE1 </td> <td> mCherry was first PCR amplified from pMZ300 with oligos oMZ701/oMZ706, further amplified with oligos oMZ703/oMZ707, and Gibson-cloned into pMZ331 cut with XbaI/EcoRI </td> </tr> | ||
<tr> <td> pIG4001 </td> <td> PhyB (1-908) - mCherry- sV40pA </td> <td> </td> <td> bla,ColE1 </td> <td> mCherry was first PCR amplified from pMZ300 with oligos oMZ701/oMZ706, further amplified using the oligos oMZ704/oMZ707 and Gibson-cloned into pMZ358 cut with XbaI/EcoRI. </td> </tr> | <tr> <td> pIG4001 </td> <td> PhyB (1-908) - mCherry- sV40pA </td> <td> </td> <td> bla,ColE1 </td> <td> mCherry was first PCR amplified from pMZ300 with oligos oMZ701/oMZ706, further amplified using the oligos oMZ704/oMZ707 and Gibson-cloned into pMZ358 cut with XbaI/EcoRI. </td> </tr> | ||
| - | <tr> <td> pIG4002 </td> <td> hU6 - precrRNA-site - CAG - | + | <tr> <td> pIG4002 </td> <td> hU6 - precrRNA-site - CAG - dCas9, D10A </td> <td> </td> <td> AmpR </td> <td> Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong et al (Science. 2013 Jan 3. PubMed) </td> </tr> |
<tr> <td> pIG4003 </td> <td> PIF3- mEGFP-sV40pA </td> <td> </td> <td> bla,ColE1 </td> <td> mEGFP was first PCR amplified from pMZ1050 with oligos oMZ728/oMZ729 and backbone pMZ354 was PCR amplified with oMZ726/oMZ727 for Gibson-Cloning. </td> </tr> | <tr> <td> pIG4003 </td> <td> PIF3- mEGFP-sV40pA </td> <td> </td> <td> bla,ColE1 </td> <td> mEGFP was first PCR amplified from pMZ1050 with oligos oMZ728/oMZ729 and backbone pMZ354 was PCR amplified with oMZ726/oMZ727 for Gibson-Cloning. </td> </tr> | ||
| - | <tr> <td> pIG4004 </td> <td> PhyB(1-605)- mCherry- | + | <tr> <td> pIG4004 </td> <td> PhyB(1-605)- mCherry- dCas9-sV40pA </td> <td> pMZ700 </td> <td> bla,ColE1 </td> <td> mCherry was amplified from pMZ700 with oligos oHB058/ oHB059, dCas9 (hSpCsn1) was amplified from pMZ1025 with oHB060/ oHB061 and Gibson- Cloned into pMZ700 (after digestion of with XbaI/ EcorI </td> </tr> |
| - | <tr> <td> pIG4005 </td> <td> PhyB(1-908)- mCherry- | + | <tr> <td> pIG4005 </td> <td> PhyB(1-908)- mCherry- dCas9-sV40pA </td> <td> pMZ701 </td> <td> bla,ColE1 </td> <td> mCherry was amplified from pIG4001 with oligos oIG4004/ oIG4001, dCas9 (hSpCsn1) was amplified from pHB1025 with oIG4002/ oIG4003 and Gibson- Cloned into pMZ701 (after digestion of with XbaI/ EcorI </td> </tr> |
| - | <tr> <td> pIG4006 </td> <td> PhyB(1-908)- mCherry- | + | <tr> <td> pIG4006 </td> <td> PhyB(1-908)- mCherry- dCas9 (with NLS)-sV40pA </td> <td> pMZ701 </td> <td> bla,ColE1 </td> <td> mCherry was amplified from pIG4001 with oligos oIG4004/ oIG4001, dCas9 (hSpCsn1) was amplified from pIG4002 with oIG4002/ oIG4005 (+NLS!) and Gibson- Cloned into pIG4001(after digestion of pIG4001 with XbaI/ EcorI) </td> </tr> |
| - | <tr> <td> pIG4007 </td> <td> PhyB(1-650)- mCherry- | + | <tr> <td> pIG4007 </td> <td> PhyB(1-650)- mCherry- dCas9 (with NLS)-sV40pA </td> <td> pMZ701 </td> <td> bla,ColE1 </td> <td> mCherry was amplified from pIG4000 with oligos oIG4000/ oIG4001, dCas9 (hSpCsn1) was amplified from pHB1025 with oIG4002/ oIG4005 and Gibson- Cloned into pIG4000 (after digestion of pIG4000 with XbaI/ EcorI </td> </tr> |
| Line 454: | Line 478: | ||
<tr> <td> pIG6000 </td> <td> pSb1C3_-Nhe1-pCMV-Sac2-eGFP-SalI-BanH1-NLS-terminator-Hind3-Kpn1-pSb1C3 </td> <td> </td> <td> </td> <td> CMV promotor was amplified from Plasmid golgi-Gfp 405(kindly provided by toolbox) using oIG6000 and oIG6001. Avgfp was amplified from Plasmid golgi-Gfp 405 using oIG6002 and oIG6003. (iGEM cutting site was eliminated) BamH1 and Sal1were introduced to provide restrcitionsites for the insertion of caspase cleavage sites. a NLS was introduced. the terminator was amplified from Plasmid pmh 105 using oIG6022 and oIG6023. thereby Kpn1 und Nhe1 and the iGEM restrcitionsites Spe1 and Not1 were reestablished. Plasmid pSB1C3 serving as backbone was cut with Xba1 and Pst1. The 4 construts were fused together with a Gibson Assemby approach. </td> </tr> | <tr> <td> pIG6000 </td> <td> pSb1C3_-Nhe1-pCMV-Sac2-eGFP-SalI-BanH1-NLS-terminator-Hind3-Kpn1-pSb1C3 </td> <td> </td> <td> </td> <td> CMV promotor was amplified from Plasmid golgi-Gfp 405(kindly provided by toolbox) using oIG6000 and oIG6001. Avgfp was amplified from Plasmid golgi-Gfp 405 using oIG6002 and oIG6003. (iGEM cutting site was eliminated) BamH1 and Sal1were introduced to provide restrcitionsites for the insertion of caspase cleavage sites. a NLS was introduced. the terminator was amplified from Plasmid pmh 105 using oIG6022 and oIG6023. thereby Kpn1 und Nhe1 and the iGEM restrcitionsites Spe1 and Not1 were reestablished. Plasmid pSB1C3 serving as backbone was cut with Xba1 and Pst1. The 4 construts were fused together with a Gibson Assemby approach. </td> </tr> | ||
<tr> <td> pIG6001 </td> <td> pSb1C3_-Nhe1-pCMV-Sac2-CFP-SalI-BanH1-NLS-terminator-Hind3-Kpn1-pSb1C3 </td> <td> </td> <td> </td> <td> CMV promotor was amplified from Plasmid golgi-Gfp 405(kindly provided by toolbox) using oIG6000 and oIG6001. ecfp was amplified from Plasmid pal 113 using oIG6002 and oIG6003. (iGEM cutting site was eliminated) BamH1 and Sal1were introduced to provide restrcitionsites for the insertion of caspase cleavage sites. a NLS was introduced. the terminator was amplified from Plasmid pmh 105 using oIG6022 and oIG6023. thereby Kpn1 und Nhe1 and the iGEM restrcitionsites Spe1 and Not1 were reestablished. Plasmid pSB1C3 serving as backbone was cut with Xba1 and Pst1. The 4 construts were fused together with a Gibson Assemby approach. </td> </tr> | <tr> <td> pIG6001 </td> <td> pSb1C3_-Nhe1-pCMV-Sac2-CFP-SalI-BanH1-NLS-terminator-Hind3-Kpn1-pSb1C3 </td> <td> </td> <td> </td> <td> CMV promotor was amplified from Plasmid golgi-Gfp 405(kindly provided by toolbox) using oIG6000 and oIG6001. ecfp was amplified from Plasmid pal 113 using oIG6002 and oIG6003. (iGEM cutting site was eliminated) BamH1 and Sal1were introduced to provide restrcitionsites for the insertion of caspase cleavage sites. a NLS was introduced. the terminator was amplified from Plasmid pmh 105 using oIG6022 and oIG6023. thereby Kpn1 und Nhe1 and the iGEM restrcitionsites Spe1 and Not1 were reestablished. Plasmid pSB1C3 serving as backbone was cut with Xba1 and Pst1. The 4 construts were fused together with a Gibson Assemby approach. </td> </tr> | ||
| - | <tr> <td> pIG8002 </td> <td> HA-NLS- | + | <tr> <td> pIG8002 </td> <td> HA-NLS-dCas9-GSAGSAG-G9a_NLS </td> <td> pX334a </td> <td> amp </td> <td> </td> </tr> |
<tr> <td> pIG8002_1 </td> <td> pIG8002 cut with BbsI and ligated with VEGF crRNA1 </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG8002_1 </td> <td> pIG8002 cut with BbsI and ligated with VEGF crRNA1 </td> <td> </td> <td> </td> <td> </td> </tr> | ||
<tr> <td> pIG8002_2 </td> <td> pIG8002 cut with BbsI and ligated with VEGF crRNA2 </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG8002_2 </td> <td> pIG8002 cut with BbsI and ligated with VEGF crRNA2 </td> <td> </td> <td> </td> <td> </td> </tr> | ||
<tr> <td> pIG8002_3 </td> <td> pIG8002 cut with BbsI and ligated with VEGF crRNA3 </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG8002_3 </td> <td> pIG8002 cut with BbsI and ligated with VEGF crRNA3 </td> <td> </td> <td> </td> <td> </td> </tr> | ||
<tr> <td> pIG8002_4 </td> <td> pIG8002 cut with BbsI and ligated with VEGF crRNA4 </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG8002_4 </td> <td> pIG8002 cut with BbsI and ligated with VEGF crRNA4 </td> <td> </td> <td> </td> <td> </td> </tr> | ||
| - | <tr> <td> pIG8004 </td> <td> HA-NLS- | + | <tr> <td> pIG8004 </td> <td> HA-NLS-dCas9-GSAGSAG-dG9a_NLS </td> <td> pX334a </td> <td> amp </td> <td> </td> </tr> |
<tr> <td> pIG8004_1 </td> <td> pIG8004 cut with BbsI and ligated with VEGF crRNA1 </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG8004_1 </td> <td> pIG8004 cut with BbsI and ligated with VEGF crRNA1 </td> <td> </td> <td> </td> <td> </td> </tr> | ||
<tr> <td> pIG8004_2 </td> <td> pIG8004 cut with BbsI and ligated with VEGF crRNA2 </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG8004_2 </td> <td> pIG8004 cut with BbsI and ligated with VEGF crRNA2 </td> <td> </td> <td> </td> <td> </td> </tr> | ||
| Line 465: | Line 489: | ||
<tr> <td> pIG8004_4 </td> <td> pIG8004 cut with BbsI and ligated with VEGF crRNA4 </td> <td> </td> <td> </td> <td> </td> </tr> | <tr> <td> pIG8004_4 </td> <td> pIG8004 cut with BbsI and ligated with VEGF crRNA4 </td> <td> </td> <td> </td> <td> </td> </tr> | ||
<tr> <td> pIG4100 </td> <td> tetO13 - CMV Promotor - SEAP - SV40 </td> <td> T405 </td> <td> Amp </td> <td> </td> </tr> | <tr> <td> pIG4100 </td> <td> tetO13 - CMV Promotor - SEAP - SV40 </td> <td> T405 </td> <td> Amp </td> <td> </td> </tr> | ||
| - | <tr> <td> pIG4101 </td> <td> pSV40 - NLS - | + | <tr> <td> pIG4101 </td> <td> pSV40 - NLS - dCas9 - NLS - Linker - PIF - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> |
<tr> <td> pIG4102 </td> <td> pSV40 - PhyB - KRAB - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> | <tr> <td> pIG4102 </td> <td> pSV40 - PhyB - KRAB - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> | ||
| - | <tr> <td> pIG4103 </td> <td> pSV40 - NLS - | + | <tr> <td> pIG4103 </td> <td> pSV40 - NLS - dCas9 - NLS - Linker - UVR8 - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> |
<tr> <td> pIG4104 </td> <td> pSV40 - COP1 - KRAB - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> | <tr> <td> pIG4104 </td> <td> pSV40 - COP1 - KRAB - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> | ||
| - | <tr> <td> pIG4105 </td> <td> pSV40 - NLS - | + | <tr> <td> pIG4105 </td> <td> pSV40 - NLS - dCas9 - NLS - Linker - CRY2 - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> |
<tr> <td> pIG4106 </td> <td> pSV40 - CIB - VP16 - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> | <tr> <td> pIG4106 </td> <td> pSV40 - CIB - VP16 - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> | ||
<tr> <td> pIG4107 </td> <td> pSV40 - CIB - KRAB - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> | <tr> <td> pIG4107 </td> <td> pSV40 - CIB - KRAB - SV40 </td> <td> </td> <td> Amp </td> <td> </td> </tr> | ||
<tr> <td> pIG4108 </td> <td> crRNA tetO13 1.CC </td> <td> </td> <td> Amp </td> <td> </td> </tr> | <tr> <td> pIG4108 </td> <td> crRNA tetO13 1.CC </td> <td> </td> <td> Amp </td> <td> </td> </tr> | ||
<tr> <td> pIG4109 </td> <td> crRNA tetO13 2.CC </td> <td> </td> <td> Amp </td> <td> </td> </tr> | <tr> <td> pIG4109 </td> <td> crRNA tetO13 2.CC </td> <td> </td> <td> Amp </td> <td> </td> </tr> | ||
| + | <tr> <td> pIG9000 </td> <td> hU6 - EMX-crRNA - HA - NLS - dCas9 - NLS </td> <td> px334a </td> <td> Amp </td> <td> </td> </tr> | ||
| + | </table> | ||
</div> | </div> | ||
</div> | </div> | ||
Latest revision as of 22:35, 4 October 2013
| | ||
|---|---|---|
| oligo name | Sequence 5'-3' | description |
| oIG0000 | TGAGCAAGAGGTAAGGG | Sequencing Primer |
| oIG0001 | AAACTGGTGGACAGCAC | Sequencing Primer |
| oIG0001_new | GGACAGCACCGACAAGG | Sequencing Primer |
| oIG0002 | AGCTGCCTGAGAAGTAC | Sequencing Primer |
| oIG0003 | TGGACCTGCTGTTCAAG | Sequencing Primer |
| oIG0004 | ACGAGCTCGTGAAAGTG | Sequencing Primer |
| oIG0005 | AGAATGACAAGCTGATCCG | Sequencing Primer |
| oIG0006 | TGGCCTATTCTGTGCTG | Sequencing Primer |
| oIG0007 | CAGGCCGAGAATATCATCC | Sequencing Primer |
| oIG0008 | CAAACAACAGATGGCTGGC | Sequencing Primer |
| oIG0009 | CTCCGATCAGGTTCTTC | Sequencing Primer |
| oIG0010 | GACAAGAAGTACAGCATCG | Sequencing Primer |
| oIG0011 | CCCGTAATGCAGAAGAAG | Sequencing Primer |
| oIG0012 | GAAGCGGAGCTACTAAC | Sequencing Primer |
| oIG0013 | GACAATGCGATGCAATTTC | Sequencing Primer |
| oIG0014 | CTTGGTCACCTTCAGCTTG | Sequencing Primer |
| oIG0015 | TTTCTTGGGTAGTTTGCAG | Sequencing Primer |
| oIG0016 | ACCAAAATCAACGGGAC | Sequencing Primer |
| oIG0017 | ACCAATGTGACCGAGTTCG | Sequencing Primer |
| oIG0018 | GCCAGCCATCTGTTGTTTG | Sequencing Primer |
| oIG0019 | AGAAGCGCGATCACATG | Sequencing Primer |
| oIG0020 | CTAATTACAGCCCGGAGGA | Sequencing Primer |
| oIG0021 | ACCAGAAAGTTAACTGG | Sequencing Primer (Bb Michael fwd) |
| oIG0022 | TGGAATTCGCGGCCGCTTCTAGATGGCCGGCGACAAGAAGTACAGCATCGG | Start+RFC25 fv |
| oIG0023 | CTGCAGCGGCCGCTACTAGTATTAACCGGTGTCGCCTCCCAGCTGAG | Ende+RFC25 rev |
| oIG0024 | TCCGGCCTTATCCAGTTCGC | 2742 (C-->A) rv |
| oIG0025 | GCGAACTGGATAAGGCCGGATTCATCAAGAGACAGCTGGTGG | 2742(C-->A) fv |
| oIG0026 | TTGCAGGTAGTACAGGTACAGCTTCTCGTTCTGAAGCTGGGTGTTTTCCACGG | 2415(G-->T)/2448 (G-->A!) rev |
| oIG0027 | TGTACCTGTACTACCTGCAAAATGGGCGGGATATGTACGTG | 2415(G-->T)/2448(G-->A!) fv |
| oIG0028 | TGGCTCCGCCGTCAATGTATCCGGCGTAGCCGTTCTTG | 1080(C-->A) rev |
| oIG0029 | ATACATTGACGGCGGAGCC | 1080(C-->A) fv |
| oIG0031 | GGGCGAACTGCAAAAGGGAAACGAACTGGCCC | 3648(C-->T)/3660(G-->A!) fv |
| oIG0032 | TTCCCTTTTGCAGTTCGCCCGCAGAGGCCAGCATTCTCTTC | 3648(C-->G)/3660(G-->A!) rev |
| oIG0033 | AGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAAACAGTGAAGGTGGTGGACGAGCTCGTGAAAGTGATGGGACGGCACAAGCCCGAGAAC | 2181 (C-->A)/2214(G-->A!)/2253(C-->A) fv |
| oIG0034 | TCTTAATGGCGGGGCTGCCTGCCAGATTGGCAATGTGCTC | 2181(C-->A)/2214(G-->A!)/2253(C-->A)rev |
| oIG0035 | TGGCCGAGGATGCCAAACTTCAGCTGAGCAAGGACACCTAC | 789 (G-->T) fv |
| oIG0036 | AGGTGTCCTTGCTCAGCTGAAGTTTGGCATCCTCGGCC | 789(G-->C) rev |
| oIG0037 | ACCGGTTAATACTAGTAGCGGCCGC | pSB1C3-RFC25_fw (AgeI, SpeI, NotI) |
| oIG0038 | GCCGGCCATCTAGAAGCGGC | pSB1C3-RFC25_rev (NgoMIV, XbaI) |
| oIG6017 | GAGTGCCACCTGACGTCTAAGAAAC | primerpBS1C3fw sequencing |
| oIG6018 | CGCCTTTGAGTGAGCTGATACCGC | primerpBS1C3rv sequencing |
| oIG0039 | ATGATTTCTGGAATTCGCGGCCGCTTCTAGATGGCCGGCGCGCGTACGAAAAACAATTACGGGTC | VP16 [25] for |
| oIG0040 | TTTTGCCGGACTGCAGCGGCCGCTACTAGTATTAACCGGTCCCACCGTACTCGTCAATTCCAAGG | VP16 [25] rev |
| oIG0041 | ATGATTTCTGGAATTCGCGGCCGCTTCTAGATGGCCGGCATGGATGCTAAGTCACTAACTGCCTGG | KRAB [25] for |
| oIG0042 | TTTTGCCGGACTGCAGCGGCCGCTACTAGTATTAACCGGTCCAGAGATCATTCCTTGCCATTCCTTC | KRAB [25] rev |
| oIG0043 | TTCTGGAATTCGCGGCCGCTTCTAGATGGCCGGCCTGTGGAATGTGTGTCAGTTAGGGTG | SV40 Promoter [25] fw |
| oIG0044 | CGGACTGCAGCGGCCGCTACTAGTATTAACCGGTAGCTTTTTGCAAAAGCCTAGGCCTCC | SV40 Promoter [25] rev |
| oIG0045 | CCGGCGCAGGCTCCACCGGTTAATA | Linker: Scar (Ala-Gly) - Ala-Gly-Ser - Scar (Thr-Gly) [25] fw |
| oIG0046 | CTAGTATTAACCGGTGGAGCCTGCG | Linker: Scar (Ala-Gly) - Ala-Gly-Ser - Scar (Thr-Gly) [25] fw |
| oIG0047 | CCGGCTACCCATACGATGTTCCAGATTACGCTACCGGTTAATA | HA-tag – fw primer |
| oIG0048 | CTAGTATTAACCGGTAGCGTAATCTGGAACATCGTATGGGTAG | HA-tag rev primer |
| oIG0049 | TGATTTCTGGAATTCGCGGCCGCTTCTAGATGGCCGGCGTGCCTTCTAGTTGCCAGCC | BGH-Terminator fv |
| oIG0050 | CCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATTAACCGGTCATAGAGCCCACCGCATCC | BGH-Terminator rev |
| oIG0051 | CCGGCGATAGTGCTGGTAGTGCTGGTAGTGCTGGTACCGGTTAATA | DSAGSAGSAG-Linker fv |
| oIG0052 | CTAGTATTAACCGGTACCAGCACTACCAGCACTACCAGCACTATCG | DSAGSAGSAG-Linker rev |
| oIG0053 | ATATCTTGTGGAAAGGACG | colony pcr pIG3010 crRNA locus for |
| oIG0054 | TTCTGCAGAATTGGCGCAC | colony pcr pIG3010 crRNA locus rev |
| oIG0055 | AACCGGTGCAGAATTGG | |
| oIG0056 | TTTTGCCGGACTGCAGCGGCCGCTACTAGTATTAACCGGTGCAGAATTGGCGCACGCGCTAG | |
| oIG0057 | AGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGGTGCCTTCTAGTTGCCAGCC | BGH (RFC 10) fwd |
| oIG0058 | CCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTACATAGAGCCCACCGCATCC | BGH (RFC 10) rev |
| oIG0059 | AGCTTTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGCTGTGGAATGTGTGTCAGTTAGGGTG | SV40 (RFC 10) fwd |
| oIG0060 | CCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTAAGCTTTTTGCAAAAGCCTAGGCCTCC | SV40 (RFC 10) rev |
| oIG1000 | CTAGCGCGTGCGCCAATTC | pCBh-fwd_pX334a |
| oIG1001 | GAAAGCTCTGAGGCACGATGGCGTCCACATCGTAGTCGGACAG | hSpCsn1-H840A-rev_pX334a |
| oIG1002 | TGTCCGACTACGATGTGGACGCCATCGTGCCTCAGAGCTTTCTG | hSpCsn-H840A-fwd_pX334a |
| oIG1003 | GTACGCGCGCGACCAGCACTACCAGCACTGTCGCCTCCCAGCTGAGAC | VP16-linker-hSpCsn-rev_pX334a |
| oIG1004 | TGGGAGGCGACAGTGCTGGTAGTGCTGGTCGCGCGCGTACGAAAAACAATTAC | hSpCsn-linker-VP16-fwd_pKM018 |
| oIG1005 | GGAGCGGGTTGATGACGTCAGCGTTCGAATTGCGGCCGCGGCAGGATCATAATCAGCCATAC | pX334a-SV40pA-rev_pKM018 |
| oIG1007 | AAACGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGT | sticky end bsbI-EMXI Locus-sticky end bsbI-fwd_pX334a |
| oIG1008 | TAAAACTTCTTCTTCTGCTCGGACTCAGGCCCTTCC | sticky end bsbI-EMXI Locus-sticky end bsbI-rev_pX334a |
| oIG1009 | CGATATCAGGAGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGGCTCTCGCGACCTGCAGGG | AatII-EcorV-emxIoperator-NruI-SbfI-NheI-fwd_pKM602 |
| oIG1010 | CTAGCCCTGCAGGTCGCGAGAGCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCCTCCTGATATCGACGT | AatII-EcorV-emxIoperator-NruI-SbfI-NheI-rev_pKM602 |
| oIG1011 | AAACTTAAAAGTCGGCTGGTAGCGGGGAGGATCGGT | human VEGFA locus 1_VZ-8_fwd_pKM600 |
| oIG1012 | TAAAACCGATCCTCCCCGCTACCAGCCGACTTTTAA | human VEGFA locus 1_VZ-8_rev_pKM600 |
| oIG1013 | AAACGTGTGCAGACGGCAGTCACTAGGGGGCGCTGT | human VEGFA locus 2_VZ-573_fwd_pKM600 |
| oIG1014 | TAAAACAGCGCCCCCTAGTGACTGCCGTCTGCACAC | human VEGFA locus 2_VZ-573_rev_pKM600 |
| oIG1015 | AAACGACCTGCTTTTGGGGGTGACCGCCGGAGCGGT | human VEGFA locus 3_VZ+434b_fwd_pKM600 |
| oIG1016 | TAAAACCGCTCCGGCGGTCACCCCCAAAAGCAGGTC | human VEGFA locus 3_VZ+434b_rev_pKM600 |
| oIG1017 | AAACGTGAGTGTGTGCGTGTGGGGTTGAGGGCGTGT | human VEGFA locus 4_VZ-475__fwd_pKM600 |
| oIG1018 | TAAAACACGCCCTCAACCCCACACGCACACACTCAC | human VEGFA locus 4_VZ-475_rev_pKM600 |
| oIG1019 | CGATATCAGGATTAAAAGTCGGCTGGTAGCGGGGAGGATCGCGGCTCTCGCGACCTGCAGGG | AatII-EcorV-VEGFA_VZ-8-NruI-SbfI-NheI-fwd |
| oIG1020 | CTAGCCCTGCAGGTCGCGAGAGCCGCGATCCTCCCCGCTACCAGCCGACTTTTAATCCTGATATCGACGT | AatII-EcorV-emxIoperator-NruI-SbfI-NheI-rev |
| oIG1021 | CGATATCAGGAGTGTGCAGACGGCAGTCACTAGGGGGCGCTCGGCTCTCGCGACCTGCAGGG | AatII-EcorV-VEGFA_ VZ-573-NruI-SbfI-NheI-fwd |
| oIG1022 | CTAGCCCTGCAGGTCGCGAGAGCCGAGCGCCCCCTAGTGACTGCCGTCTGCACACTCCTGATATCGACGT | AatII-EcorV-VEGFA_ VZ-573-NruI-SbfI-NheI-rev |
| oIG1023 | CGATATCAGGAGACCTGCTTTTGGGGGTGACCGCCGGAGCGCGGCTCTCGCGACCTGCAGGG | AatII-EcorV-VEGFA_ VZ+434b-NruI-SbfI-NheI-fwd |
| oIG1024 | CTAGCCCTGCAGGTCGCGAGAGCCGCGCTCCGGCGGTCACCCCCAAAAGCAGGTCTCCTGATATCGACGT | AatII-EcorV-VEGFA_ VZ+434b-NruI-SbfI-NheI-rev |
| oIG1025 | CGATATCAGGAGTGAGTGTGTGCGTGTGGGGTTGAGGGCGTAGGCTCTCGCGACCTGCAGGG | AatII-EcorV-VEGFA_ VZ-475-NruI-SbfI-NheI-fwd |
| oIG1026 | CTAGCCCTGCAGGTCGCGAGAGCCTACGCCCTCAACCCCACACGCACACACTCACTCCTGATATCGACGT | AatII-EcorV-VEGFA_ VZ-475-NruI-SbfI-NheI-rev |
| oIG1027 | GTTCTCCTCCTCAACTG | Sequencing Primer pKM602 |
| oIG1028 | GTAACTTGAAAGTATTTCGATTTCTTG | DR-fwd_pX334a |
| oIG1029 | GCGGCCGCAATTCGAACG | H1Promotor-fwd_BBpX334a |
| oIG1030 | CACAAGATATATAAAGCCAAGAAATC | U6 Promotor-rev_BBpX334a |
| oIG2000 | CAGACCCCGTAGAAAAGATCAAAGGATC | Ori1-hU6-CAGenhancer-hSpdCas9-fwd_pX334a |
| oIG2001 | GAAAGCTCTGAGGCACGATAGCGTCCACATCGTAGTCGGAC | hSpdCas9-H840A-rev_pX334a |
| oIG2002 | GTCCGACTACGATGTGGACGCTATCGTGCCTCAGAGCTTTC | H840A-linkerGSAGSAG-KRAB-fwd_pX334a |
| oIG2003 | CTTAGCATCCATGCCGGCGGAGCCTGCTGATCCGGGGCTGTCGCCTCCCAGC | H840A-linkerGSAGSAG-KRAB-rev_pX334a |
| oIG2004 | GACAGCCCCGGATCAGCAGGCTCCGCCGGCATGGATGCTAAGTCACTAACTGCCTGGTC | linkerGSAGSAG-KRAB-bgHTerminator-fwd_pKM102 |
| oIG2005 | AGAAGGCACAGTCGAGGCTGATCGAAGCTTGGATCCTCACACCTTCC | linkerGSAGSAG-KRAB-bgHTerminator-rev_pKM102/pX334a |
| oIG2006 | GGAAGGTGTGAGGATCCAAGCTTCGATCAGCCTCGACTGTGCCTTCTAG | bgHTerminator-PuroR/Backbone1-fwd_pKM102/pX334a |
| oIG2007 | CACACCCGCCGCGCTTAATGCG | bgHTerminator-PuroR/Backbone1-rev_pX334a |
| oIG2008 | GCCCTGTAGCGGCGCATTAAGCG | Backbone1-AmpR-Ori1-fwd_pX334a |
| oIG2009 | GGATCTCAAGAAGATCCTTTGATCTTTTCTAC | Backbone1-AmpR-Ori1-rev_pX334a |
| oIG2010 | AAACGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGT | sticky end BbsI-EMX1-sticky end BbsI-fwd_pX334 |
| oIG2011 | TAAAACTTCTTCTTCTGCTCGGACTCAGGCCCTTCC | sticky end BbsI-EMX1-sticky end BbsI-rev_pX334 |
| oIG2012 | CTAGGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGG | sticky end NheI-EMX1-sticky end NheI-fwd_pKM006 |
| oIG2013 | CTAGCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCC | sticky end NheI-EMX1-sticky end NheI-rev_pKM006 |
| oIG2014 | AATTGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGG | sticky end EcoRI-EMX1-sticky end EcoRI-fwd_pKM006 |
| oIG2015 | AATTCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCC | sticky end EcoRI-EMX1-sticky end EcoRI-rev_pKM006 |
| oIG2016 | AAACGGATCTGCACCACCGGCAAGCTGCCCGTGCCCGT | sticky end BbsI-pKM006 target_nt650-sticky end BbsI-fwd_pX334 |
| oIG2017 | TAAAACGGGCACGGGCAGCTTGCCGGTGGTGCAGATCC | sticky end BbsI-pKM006 target_nt650-sticky end BbsI-rev_pX334 |
| oIG2018 | AAACGGCACGACTTCTTCAAGTCCGCCATGCCCGAGT | sticky end BbsI-pKM006 target_nt750-sticky end BbsI-fwd_pX334 |
| oIG2019 | TAAAACTCGGGCATGGCGGACTTGAAGAAGTCGTGCC | sticky end BbsI-pKM006 target_nt750-sticky end BbsI-rev_pX334 |
| oIG2020 | AAACGGCGACACCCTGGTGAACCGCATCGAGCTGAAGT | sticky end BbsI-pKM006 target_nt850-sticky end BbsI-fwd_pX334 |
| oIG2021 | TAAAACTTCAGCTCGATGCGGTTCACCAGGGTGTCGCC | sticky end BbsI-pKM006 target_nt850-sticky end BbsI-rev_pX334 |
| oIG2022 | AAACGGCGTCTATATCACCGCCGACAAGCAGAAGAAGT | sticky end BbsI-pKM006 target_nt950-sticky end BbsI-fwd_pX334 |
| oIG2023 | TAAAACTTCTTCTGCTTGTCGGCGGTGATATAGACGCC | sticky end BbsI-pKM006 target_nt950-sticky end BbsI-rev_pX334 |
| oIG2024 | AAACGGAGAGCTCGTTTAGTGAACCGTCAGATCGCCGT | sticky end BbsI-pKM006 target_CMVmin1-sticky end BbsI-fwd_pX334 |
| oIG2025 | TAAAACGGCGATCTGACGGTTCACTAAACGAGCTCTCC | sticky end BbsI-pKM006 target_CMVmin1-sticky end BbsI-rev_pX334 |
| oIG2026 | AAACGGCACGCTGTTTTGACCTCCATAGAAGACACCGT | sticky end BbsI-pKM006 target_CMVmin2-sticky end BbsI-fwd_pX334 |
| oIG2027 | TAAAACGGTGTCTTCTATGGAGGTCAAAACAGCGTGCC | sticky end BbsI-pKM006 target_CMVmin2-sticky end BbsI-rev_pX334 |
| oIG2028 | CCGGTGCCACCATGTACCCATAC | hSpdCas9-H840A-fwd_pX334a |
| oIG2029 | CGTAATCTGGAACATCGTATGGGTACATG | Ori1-hU6-CAGenhancer-hSpdCas9-H840A-rev_pX334a |
| oIG2030 | AAACCAAGGTCGGAGTACTGTCCTCCGACACTAGGT | sticky end bsbI-UAS_Zebrafish_Locus_pos58-sticky end bsbI-fwd |
| oIG2031 | TAAAACCTAGTGTCGGAGGACAGTACTCCGACCTTG | sticky end bsbI-UAS_Zebrafish_Locus_pos58-sticky end bsbI-rev |
| oIG2032 | AAACGAAATTCGGTCCATCCGCGCGCGCAGAGTGGT | sticky end bsbI-Bmp2b_Zebrafish_Locus_pos55-sticky end bsbI-fwd |
| oIG2033 | TAAAACCACTCTGCGCGCGCGGATGGACCGAATTTC | sticky end bsbI-Bmp2b_Zebrafish_Locus_pos55-sticky end bsbI-rev |
| oIG2034 | AAACAACTCCTCCATGCATATGTTAATGCGCCGCGT | sticky end bsbI-Pouf51-VOX_Zebrafish_Locus_pos125-sticky end bsbI-fwd |
| oIG2035 | TAAAACGCGGCGCATTAACATATGCATGGAGGAGTT | sticky end bsbI-Pouf51-VOX_Zebrafish_Locus_pos125-sticky end bsbI-rev |
| oIG2036 | TAATACGACTCACTATAGGGACGAAACACCGGGTTTTAGAGC | crRNA with T7 site (forward) |
| oIG2037 | CACGCGCTAGAAAAAGTTTTGGGACC | crRNA (reverse) |
| oIG2038 | TAATACGACTCACTATAGGGACCGGTGCCACCATGTACC | dCas9 with T7 site and Kozak-sequence (forward) |
| oIG2039 | AATTGCGGCCGCTCCCCAGC | bGH Terminator (reverse) |
| oIG2040 | GCAATAGCATCACAAATTTCACAAATAAAGC | SV40 Terminator (reverse) |
| oIG2041 | TAATACGACTCACTATAGGGGTTGGAACCATTCAAAACAGCATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTGGCG | tracrRNA with T7 site (forward) |
| oIG2042 | CGCCAAAAAGCACCGACTCGGTGCCACTTTTTCAAGTTGATAACGGACTAGCCTTATTTTAACTTGCTATGCTGTTTTGAATGGTTCCAACCCCTATAGTGAGTCGTATTA | tracrRNA (reverse) |
| oIG2043 | AAACGATGCGGTTTTGGCAGTACATCAATGGGCGGT | sticky end BbsI-CMV target of pIG6000-sticky end BbsI-fwd |
| oIG2044 | TAAAACCGCCCATTGATGTACTGCCAAAACCGCATC | sticky end BbsI-CMV target of pIG6000-sticky end BbsI-rev |
| oIG3001 | GGCAGCGCCGGCTCCGGCAGCGCCTCCGGCATGGTGAGCAAGGGCGAGGAGG | Linker1-mCherry_forw_pMZ333 |
| oIG3002 | TCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATCACTTGTACAGCTCGTCCATGCCGC | mCherry-Bb_rev_pMZ333 |
| oIG3003 | TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGCTGACAAGAAGTACAGCATCGGCCTG | Bb-HA-dCas9_forw_pX334a |
| oIG3004 | GCCGGAGGCGCTGCCGGAGCCGGCGCTGCCGTCGCCTCCCAGCTGAGACAG | dCas9-Linker1_rev_pX334a |
| oIG3005 | TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGC | Bb-HA(-NLS-dCas9)_forw_pMZ333 |
| oIG3006 | TCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATCAGCTGGCCTCCACTTTTCTCTTCTTTTTGGGGCTCTTGTACAGCTCGTCCATGCCGC | mCherry-NLS-Bb_rev_pMZ333 |
| oIG3007 | TGTCTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCAGGAGGCGCCACCATGTACCCATACGATGTTCCAGATTACGCTATGAGAGCTGCCAACCTTTGGCCAAG | Bb-HA-ERT2_forw_pMItom |
| oIG3008 | GGACGCTTCGACCTTGCGCTTTTTCTTCGGCGAGACTGTGGCAGGGAAACCCTCTG | ERT2-NLS(-dCas9)_rev_pMItom |
| oIG3009 | GGGGAGGCAGAGGGTTTCCCTGCCACAGTCTCGCCGAAGAAAAAGCGCAAGGTC | ERT2-NLS(-dCas9)_forw_pX334a |
| oIG3010 | GCTGGCCTCCACTTTTCTCTTCTTTTTGGGGCTCTTGTACAGCTCGTCCATGCCGC | mCherry-NLS3'_rev_pMZ333 |
| oIG3011 | AGCCCCAAAAAGAAGAGAAAAGTGGAGGCCAGCATGAGAGCTGCCAACCTTTGGCCAAG | NLS3'-ERT2_forw_pMItom |
| oIG3012 | TCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATCAGACTGTGGCAGGGAAACCCTCTG | ERT2-Bb_rev_pMItom |
| oIG3013 | AAACGATGATGAGGTTCTTGGCGGCTGTCTGTGCGT | crRNA_SEAP(nt136-166)_forw |
| oIG3014 | TAAAACGCACAGACAGCCGCCAAGAACCTCATCATC | crRNA_SEAP(nt136-166)_rev |
| oIG3015 | AAACCTGGCACATGTTTGTCTACATTGTATGTCTGT | crRNA_SEAP(nt293-323)_forw |
| oIG3016 | TAAAACAGACATACAATGTAGACAAACATGTGCCAG | crRNA_SEAP(nt293-323)_rev |
| oIG4000 | GATATGGCGGGGGAACAGGGG | mCherry-fwd_pMZ700 |
| oIG4001 | GCCGCTGCCGGCGCCGCTGCCGGCGCTGCCCTTGTACAGCTCGTCCATGCCGC | mCherry-rev_pMZ700/pMZ701 |
| oIG4002 | GGCAGCGCCGGCAGCGGCGCCGGCAGCGGCGACAAGAAGTACAGCATCGGCCTG | dCas9-fwd_pMZ1025 |
| oIG4003 | GTCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATTAGTCGCCTCCCAGCTGAGACAG | dCas9-rev_pMZ1025 |
| oIG4004 | GTTATTGGGGCTTTCTGTTTCTTGCAAATCC | mCherry-fwd_pMU701 |
| oIG4005 | GTCTGGATCGAAGCTTGGGCTGCAGGTCGACTCTAGATTACACCTTTCTCTTCTTCTTGGGGCTGTC | dCas9(with NLS)-rev_pMZ1025 |
| oIG4006 | TCGAAGCTTGGGCTGCAGGTCGACTCTAGATTACACCTTTCTCTTCTTCTTGGGGCTGTCGCCTCCCAGCTGAGACAG | dCas9(with NLS)-rev_pIG4004 and pIG4005 |
| oIG5000 | TGCCTGAAATCACTTTTTTTCAGGTTGGACCGGTGCCACCTCGCGAATGTACCCATACGATGTTCCAGATTACG | Backbone-Nru1-HA_for_pX334a |
| oIG5001 | GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCATGGCCGACAAGCAGAAGAACGGC | (GGGGS)3_Linker-BamH1-splitC_YFP_for_BBa_K323080 |
| oIG5002 | AAGTTAGTAGCTCCGCTTCCGCTGGCCTCCACCTTTCTCTTCTTCTTGGGGCTGTCGACCTTGTACAGCTCGTCCATGCCGAG | P2A(first 20bp)-NLS-Sal1-splitC_YFP_rev_BBa_K323080 |
| oIG5003 | GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCGTGAGCAAGGGCGAGGAGCTG | (GGGGS)3_Linker-BamH1-splitN_YFP_for_BBa_K323005 |
| oIG5004 | AAGTTAGTAGCTCCGCTTCCGCTGGCCTCCACCTTTCTCTTCTTCTTGGGGCTGTCGACGATGTTGTGGCGGATCTTGAAGTTCAC | P2A(first20 bp)-NLS-Sal1-splitN_YFP_rev_BBa_K323005 |
| oIG5007 | GGAAGCGGAGCTACTAACTTCAGCCTGCTGAAGCAGGCTGGAGACGTGGAGGAGAACCCTGGACCTATGGTGAGCAAGGGCGAGGAGG | P2A-mCherry_for_pMH206 |
| oIG5008 | CAACTAGAAGGCACAGTCGAGGCTGATCAGCGAGCTCTAGTCGCGATTACTTGTACAGCTCGTCCATGCCGC | Backbone(pX334a)-Nru1-mCherry_rev_pMH206 |
| oIG5009 | CTAGAGCTCGCTGATCAGCCTCG | forward - pX334a backbone amplification (Part1) |
| oIG5010 | TCAAAAGAATAGCCCGAGATAGGGTTGAG | reverse - pX334a backbone amplification (Part1) |
| oIG5011 | CTTTAATAGTGGACTCTTGTTCCAAACTGGAAC | forward - pX334a backbone amplification (Part2) |
| oIG5012 | GGTGGCACCGGTCCAACCTG | reverse - pX334a backbone amplification (Part2) |
| oIG5013 | ACTGCCTCCGCCACCACTGCCACCGCCTCCGTCGCCTCCCAGCTGAGACAG | (GGGGS)3_Linker-dCas9_rev_pX334a |
| oIG5014 | GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCATGGAGAACACTGAAAACTCAGTGGATTC | (GGGGS)3_Linker-BamH1-Caspase3_for_pMK31 |
| oIG5015 | AAGTTAGTAGCTCCGCTTCCGCTGGCCTCCACCTTTCTCTTCTTCTTGGGGCTGTCGACATGATGATGATGATGATGGTGATAAAAATAGAGTTC | P2A(first 20bp)-NLS-Sal1-Caspase3_rev_pMK31 |
| oIG5016 | CTTAGCGGCAGCTTCCTTAGCGGCAGCTTCGTCGCCTCCCAGCTGAGACAG | (EAAAK)3_Linker-dCas9_rev_pX334a |
| oIG5017 | GAAGCTGCCGCTAAGGAAGCTGCCGCTAAGGGATCCATGGAGAACACTGAAAACTCAGTGGATTCAAAATC | (EAAAK)3_Linker-BamH1-Caspase3_for_pMK31 |
| oIG5018 | GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGGATCCAGGCAAGCAGCAAAGTTGTCGAAGCC | (GGGGS)3_Linker-BamH1-Casp9_for_pKJ42 |
| oIG5019 | AAGTTAGTAGCTCCGCTTCCGCTGGCCTCCACCTTTCTCTTCTTCTTGGGGCTGTCGACTGATGTTTTAAAGAAAAGTTTTTTCCGGAGG | P2A(first 20bp)-NLS-sal1-Casp9(long)_rev_pKJ42 |
| oIG5021 | GAAGCTGCCGCTAAGGAAGCTGCCGCTAAGGGATCCAGGCAAGCAGCAAAGTTGTCGAAGCC | (EAAAK)2-BamH1-Casp9_for_pKJ42 |
| oIG5022 | GTCTATATAAGCAGAGCTCTCTGGCATGAGCCCCAAGAAGAAGAGAAAGGTGGAGGCCAGCATGTCGAATTCTTATGACTCCTCCAGTATC | Backbone(pcDNA_3.1)-NLS-GyrB-Caspase3_for_pMK31 |
| oIG5023 | GTCTATATAAGCAGAGCTCTCTGGCATGAGCCCCAAGAAGAAGAGAAAGGTGGAGGCCAGCAGGCAAGCAGCAAAGTTGTCGAAGC | Backbone(pcDNA_3.1)-NLS-Caspase9-GyrB_for_pKJ42 |
| oIG5024 | AAGTTAGTAGCTCCGCTTCCGCTGGCCTCCACCTTTCTCTTCTTCTTGGGGCTGCCTTCATAGTGGAAGTGGTCTTCTTTG | P2A(first 20bp)-NLS-Caspase9-GyrB_rev_pKJ42 |
| oIG5025 | ACTGCCTCCGCCACCACTGCCACCGCCTCCGCCTTCATAGTGGAAGTGGTCTTCTTTG | (GGGGS)2_Linker-GyrB_rev_pMK31 |
| oIG5026 | ATGGCTGGCAACTAGAAGGCACAGTCGAGGTCACTTGTACAGCTCGTCCATGCCGC | Backbone(pcDNA_3.1)-mCherry_rev_pMH206 |
| oIG5027 | CCTCGACTGTGCCTTCTAGTTGCC | forward - pcDNA_3.1 backbone amplification (Part1) |
| oIG5028 | CACTTTATGCTTCCGGCTCGTATGTTG | reverse - pcDNA_3.1 backbone amplification (Part1) |
| oIG5029 | CCACACAACATACGAGCCGGAAGC | forward - pcDNA_3.1 backbone amplification (Part2) |
| oIG5030 | GCCAGAGAGCTCTGCTTATATAGACC | reverse - pcDNA_3.1 backbone amplification (Part2) |
| oIG5031 | CAGGTTGGACCGGTGCCACCTCGCGAATGAGCCCCAAGAAGAAGAGAAAGGTGGAGGCCAGCGGATCCAGGCAAGCAGCAAAGTTGTCGAAGC | Backbone(pX334a)-Nru1-NLS-BamH1-Caspase9(long)_for_pKJ42 |
| oIG5032 | ACTGCCTCCGCCACCACTGCCACCGCCTCCGTCGACTGATGTTTTAAAGAAAAGTTTTTTCCGGAGG | (GGGGS)2_Linker_Sal1_Caspase9(long)_rev_pKJ42 |
| oIG5033 | ||
| oIG5034 | GGAGGCGGTGGCAGTGGTGGCGGAGGCAGTGACAAGAAGTACAGCATCGGCCTGG | (GGGGS)2_Linker-dCas9-NLS_for_pX334a |
| oIG5035 | CTTCAGCAGGCTGAAGTTAGTAGCTCCGCTTCCAGCGTAATCTGGAACATCGTATGGGTACATGCTGGCCTCCACCTTTCTCTTCTTC | P2A(first 33bp)-HA-dCas9-NLS_rev_pX334a |
| oIG6000 | TCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGCAGCTAGCTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTC | pCMVfor-psB1C3 |
| oIG6001 | CGAGCGGTCCGCGGAACCAGCGCTACCGGACTGCCATTGCTCCGGCTGATCTGACGGTTCACTAAACCAGCTC | pCMVrev-eGFP |
| oIG6002 | AGCCGGAGCAATGGCAGTCCGGTAGCGCTGGTTCCGCGGACCGCTCGGCCACCATGGTGAGCAAGGGC | eGFPfor-CMV |
| oIG6003 | GGACGCTTCGACCTTGCGCTTTTTCTTCGGCGAGGGATCCGACCCGTCGACGCCCTTGTACAGCTCGTCCATGC | eGFPrev-Sal1-Bamh1-NLS |
| oIG6004 | TCGCCGAAGAAAAAGCGCAAGGTCGAAGCGTCCTAAAGATCTATAATCAGCCATACCACATTTGTAGAGGTTTTAC | terminatorfor-NLS |
| oIG6005 | GCCGGACTGCAGCGGCCGCTACTAGTAAGGTACCGTAAGCTTTAGCTCTGAATGCATGCGTATTGCTAATGGGTCTAATGCAGTTAAGATACATTGATGAGTTTGGACAAACCAC | terminatorrev-Nru1-Hind3-pSB1C3-Spe1-Not1-Pst1 |
| oIG6006 | AAACCCCTCGCCCTCGCCGGACACGCTGAACTTGGT | eGFPfw |
| oIG6007 | TAAAACCAAGTTCAGCGTGTCCGGCGAGGGCGAGGG | eGFPrv |
| oIG6008 | AAACGGCGGGCCATTTACCGTAAGTTATGTAACGGT | CMVfw |
| oIG6009 | TAAAACCGTTACATAACTTACGGTAAATGGCCCGCC | CMVrv |
| oIG6010 | AAACTTATGTTTCAGGTTCAGGGGGAGGTGTGGGGT | sv40 terminatorfw |
| oIG6011 | TAAAACCCCACACCTCCCCCTGAACCTGAAACATAA | sv 40 terminatorrv |
| oIG6012 | GGACGCTTCGACCTTGCGCTTTTTCTTCGGCGAGGGATCCGACCCGTCGACGCCTTACTTGTACAGCTCGTCCATGCCG | eCFPrv |
| oIG605neu | GTTCCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTAAGGTACCGTAAGCTTTAGCTCTGAATGCATGCGTATTGCTAATGGGTCTAATGCAGTTAAGATACATTGATGAGTTTGGACAAACCAC | rv primer für den terminator mit mehr basen overlapp |
| oIG6015 | ATGTACACTAAGTCTAGCTAGACTAGCTGGTACCGACTAGTAGCGGCCGCTGCAGTC | |
| oIG6016 | GCTATAGCTAAGCGCGTCGAGTCGAGCGAAGCTTATCTAGAAGCGGCCGCGAATTCCAG | |
| oIG6017 | GAGTGCCACCTGACGTCTAAGAAAC | primer für pSB1C3 fw |
| oIG6018 | CGCCTTTGAGTGAGCTGATACCGC | primer für pSB1C3 rv |
| oig6020 | TCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGCAGCTAGCCGATGTACGGGCCAGATATACGCG | CMV fw |
| oig6021 | CGAGCGGTCCGCGGAACCAGCGCTACCGGACTGCCATTGCTCCGGCTAATTTCGATAAGCCAGTAAGCAGTGGGTTC | CMV rv |
| oig6022 | TCGCCGAAGAAAAAGCGCAAGGTCGAAGCGTCCTAAAGATCTCGACTGTGCCTTCTAGTTGCCAGC | bgh terminator fw |
| oig6023 | GCCGGACTGCAGCGGCCGCTACTAGTAAGGTACCGTAAGCTTTAGCTCTGAATGCATGCGTATTGCTAATGGGTCTAATGCACCATAGAGCCCACCGCATCCC | bgh terminator rv |
| oig6024 | AGCCGGAGCAATGGCAGTCCGGTAGCGCTGGTTCCGCGGACCGCTCGGCCACCATGGCCACAACCATGGTGAGCAAGG | eCFP fw |
| oig6025 | GGACGCTTCGACCTTGCGCTTTTTCTTCGGCGAGGGATCCGACCCGTCGACGCCCTTGTACAGCTCGTCCATGCCGAG | eCFP rv |
| oig6030 | AAACGTTCAAGTCGCACTCGCCGCTCCCGCTCCCGT | AvGFP fw beginning |
| oig6031 | TAAAACGGGAGCGGGAGCGGCGAGTGCGACTTGAAC | AvGFP rv beginning |
| oig6032 | AAACGTAACTGCGTTTACCCGCCATCCGCACATGGT | target CMV promoter before tata fw |
| oig6033 | TAAAACCATGTGCGGATGGCGGGTAAACGCAGTTAC | target CMV promoter before tata rv |
| oig6034 | AAACAGTCTAGTCGGCCTCGTTACCGTCAGGCCAGT | target CMV behind tata fw |
| oig6035 | TAAAACTGGCCTGACGGTAACGAGGCCGACTAGACT | target CMV behind tata rv |
| oig6036 | AAACAGATCAACGGTCGGTAGACAACAAACGGGGGT | target bgh terminator beginning fw |
| oig6037 | TAAAACCCCCGTTTGTTGTCTACCGACCGTTGATCT | target bgh terminator beginning rv |
| oig6038 | AAACGTTCAAGTCGCACAGGCCGCTCCCGCTCCCGT | target eCFP fw |
| oig6039 | TAAAACGGGAGCGGGAGCGGCCTGTGCGACTTGAAC | target eCFP rv |
| oIG8002 | GCCGGCGGAGCCTGCTGATCCGGGGCTGTCGCCTCCCAGC | dCas9_GSAGSAG_rev |
| oIG8005 | GGACGCTTCGACCTTGCGCTTTTTC | NLS1_pX334_rev |
| oIG8006 | GGATCAGCAGGCTCCGCCGGCGACAAGAAGTACAGCATCGGCCTG | dCas9_GSAGSAG_fw |
| oIG8007 | GGTGCTGGTATGACAAG | G9a_seq_fw_pGEX2T |
| oIG8008 | GATGTTGCCATAGTAACGG | G9a_seq_rev_pGEX2T |
| oIG8009 | GACAGCCCCGGATCAGCAGGCTCCGCCGGCGAGAATGTACCCATCCCCTGTGTC | G9a(mm)_C-term_GSAGSAG_fw |
| oIG8010 | GGATCCTCACACCTTCCGCTTTTTCTTGGGGGTGTTGATGGGGGGCAGGGA | G9a(mm)_C-term_NLS_bgh_rev |
| oIG8011 | AAGCGCAAGGTCGAAGCGTCCGAGAATGTACCCATCCCCTGTGTC | G9a(mm)_N-term_NLS_fw |
| oIG8012 | GCCGGCGGAGCCTGCTGATCCGGTGTTGATGGGGGGCAGGGA | G9a(mm)_N-term_GSAGSAG_rev |
| oIG8013 | GCCCGTTACTATGGCAACATCAGCCGATTCATTAACAAGCTGTGTGACCCCAACATCATCCCTGTCCGGGTTTTCATGCTG | dG9a(mm)_H->K_fw |
| oIG8014 | CAGCATGAAAACCCGGACAGGGATGATGTTGGGGTCACACAGCTTGTTAATGAATCGGCTGATGTTGCCATAGTAACGGGC | dG9a(mm)_H->K_rev |
| oIG8015 | GAATTCGCGGCCGCTTCTAGATGGCCGGCGAGAATGTACCCATCCCCTGTGTC | _G9a_fw_std |
| oIG8016 | CTGCAGCGGCCGCTACTAGTATTAACCGGTGGTGTTGATGGGGGGCAGGGA | G9a_rev_std |
| oIG8017 | GCAGTCATCCACACACGTGCAGTGCTGGAGATGGGTGATGTTGCGGTCGA | G9a_1stsite_std_rev |
| oIG8018 | CAGCACTGCACGTGTGTGGATG | G9a_1stsite_std_fw |
| oIG8019 | GCCCATCTTGGCAGTCCGGTAGAGCTGGAGCCGTACCTTGATGCCGCTCT | G9a_2ndsite_std_rev |
| oIG8020 | CAGCTCTACCGGACTGCCAAGA | G9a_2ndsite_std_fw |
| oIG8021 | CCCAAGAAAAAGCGGAAGGTGTGATC | phyB hole vector for |
| oIG8022 | GCGGCTGTACGCGGAACCAG | phyB hole vector rev |
| oIG8023 | GCTGGTAGTGCTGGTTCCGCGTACAGCCGCGAGAATGTACCCATCCCCTGTGTC | phyB(overhang)_g9a for |
| oIG8024 | CTCTAGATCACACCTTCCGCTTTTTCTTGGGGGTGTTGATGGGGGGCAGGGA | phyB(overhang)_g9a rev |
| oIG9100 | TCATATCCGCGGCATGTACCCATACGATGTTCCAGATTACG | dCas9 fw incl. HA tag and NLS |
| oIG9191 | GATATTGGTACCCATGCCTGCTATTCTCTTCCCAATCCTC | dCas9 rv incl. Bgh terminator |
| oIG4100F | GCTAGCTAGTTATTAATAGTAATCAATTACGGG | wird nachgetragen |
| oIG4100R | GAATTCGATCTGACGGTTCACTAAAC | |
| oIG4101F | ATGTACCCATACGATGTTCCAG | |
| oIG4101R | GGGGCTGTCGCCTCCCAG | |
| oIG4102F | GACACGGATCGACCTGTCTCAGCTGGGAGGCGACAGCCCCCCGAAGAAAAAGCGCAAGGTCGCGCTCGATAGTGCTGGTAG | |
| oIG4102R | TCTTCGGCGAAGCGTAATCTGGAACATCGTATGGGTACATGGTGGGCGGCCGCAATTC | |
| oIG4103F | ATGGATGCTAAGTCACTAACTGCC | |
| oIG4103R | TCACACCTTCCGCTTTTTCT | |
| oIG4104F | AAGGAATGATCTCTGGCCCAAGAAAAAGCGGAAGGTGTGATCTAGAGTCGACCTGCAGCC | |
| oIG4104R | CCAGTGTCCGGGACCAGGCAGTTAGTGACTTAGCATCCATGCGGCTGTACGCGGAACCAG | |
| oIG4105F | GACACGGATCGACCTGTCTCAGCTGGGAGGCGACAGCCCCCCGAAGAAAAAGCGCAAGGTCGGTACCGGCGCCGGCG | |
| oIG4105R | TCTTCGGCGAAGCGTAATCTGGAACATCGTATGGGTACATGGTGGGAATTCGCGGCC | |
| oIG4106F | AAGGAATGATCTCTGGCCCAAGAAAAAGCGGAAGGTGTGAGGGGCGCGAGGATCCTCTAG | |
| oIG4106R | CCAGTGTCCGGGACCAGGCAGTTAGTGACTTAGCATCCATCGCGCCGGCGCCCGCAG | |
| oIG4107F | GTCTCAGCTGGGAGGCGACAGCCCCCCGAAGAAAAAGCGCAAGGTCGGTAGTGCTGGTAGTGCTGGTAGTGCTGGTATGAAGATGGACAAAAAGACTATAGTT | |
| oIG4107R | TGCTGCTCCGATCATGATCTG | |
| oIG4108F | TTCAAGAACCCGTGAAGCACAGATCATGATCGGAGCAGCATGATCTAGAGTCGACCTGCAGC | |
| oIG4108R | TCTTCGGCGAAGCGTAATCTGGAACATCGTATGGGTACATGGTGGGCGGCCGCAATTC | |
| oIG4109F | GATAGTGCTGGTAGTGCTGGT | |
| oIG4109R | GGTGGGCGGCCGCAATTC | |
| oIG4110F | CTTTTATTTCAGGTCCCGGATCGAATTGCGGCCGCCCACCATGAATGGAGCTATAGGAGGTGAC | |
| oIG4110R | TGTACGCGGAACCAGCACTACCAGCACTACCAGCACTATCATGAATATAATCCGTTTTCTCCAATTCCT | |
| oIG4111F | AAGGAATGATCTCTGGCCCAAGAAAAAGCGGAAGGTGTGATCTAGAGTCGACCTGCAGCC | |
| oIG4111R | GGTGGGCGGCCGCAATTC | |
| oIG4112R | AGGCAGTTAGTGACTTAGCATCCATGCGGCTGTACGCGGAACCAGCACTACCAGCACTACCAGCACTATCATGAATATAATCCGTTTTCTCCAATTCCT | |
| crRNA tetO 1 F | TAAAACATCAGTGATAGAGAAAAGTGAAAGTCGAGT | |
| crRNA tetO 1 R | AAACACTCGACTTTCACTTTTCTCTATCACTGATGT | |
| crRNA tetO 2 F | TAAAACCTCCCTATCAGTGATAGAGAAAAGTGAAAG | |
| oIG9000 | AGGAGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGGCTC | ELISA CRISPR/Cas9 for EMX1 sequence, biotinylated |
| oIG9001 | GAGCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCCTCCT | ELISA CRISPR/Cas9 for EMX1 sequence |
| oIG9002 | CAGAAGCTGGAGGAGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGGCTC | ELISA CRISPR/Cas9 for EMX1 sequence, biotinylated |
| oIG9003 | GAGCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCCTCCTCCAGCTTCTG | ELISA CRISPR/Cas9 for EMX1 sequence |
| oIG9004 | CAGAAGCTGGAGGAGGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGGGCTCCCATCACATC | ELISA CRISPR/Cas9 for EMX1 sequence, biotinylated |
| oIG9005 | GATGTGATGGGAGCCCTTCTTCTTCTGCTCGGACTCAGGCCCTTCCTCCTCCAGCTTCTG | ELISA CRISPR/Cas9 for EMX1 sequence |
| oIG9006 | GGAAGGGCCTGAGTCCGAGCAGAAGAAGAAGTTTTAGAGCTATGCTGTTTTGAATGGTC | forward primer for the insertion of the EMX1 |
| oIG9007 | TTCTTCTTCTGCTCGGACTCAGGCCCTTCCGTTTTGGGACCATTCAAAACAGCATAGC | reverse primer for the insertion of the EMX1 |
| oIG9008 | GGGCTTGTACTCGGTCATGGTAAGCTTCAGCTGCTCGAGATCTAGATGGATGCAGGTCG | reverse primer for the insertion of the EMX1 |
| oIG9009 | CGACCTGCATCCATCTAGATCTCGAGCAGCTGAAGCTTACC | forward primer for the insertion of the EMX1 |
| oIG9010 | CAG CAT CGG CCT GGA CAT CGG C | Site directed mutagenesis pcr reverse primer to reestablish the endonuclease activity by activating one inactivated nicase |
| oIG9011 | GCC GAT GTC CAG GCC GAT GCT G | Site directed mutagenesis pcr reverse primer to reestablish the endonuclease activity by activating one inactivated nicase |
| oIG9012 | CAGAAGCTGGATTAAAAGTCGGCTGGTAGCGGGGAGGATCGGGGCTCCCATCACATC | human VEGFA locus 1_VZ-8_fwd_pKM600, biotinylated |
| oIG9013 | GATGTGATGGGAGCCCCGATCCTCCCCGCTACCAGCCGACTTTTAATCCAGCTTCTG | human VEGFA locus 1_VZ-8_fwd_pKM600 |
| oIG9015 | CAGAAGCTGGAGACCTGCTTTTGGGGGTGACCGCCGGAGCGGGGCTCCCATCACATC | human VEGFA locus 3_VZ+434b_fwd_pKM600, biotinylated |
| oIG9016 | GATGTGATGGGAGCCCCGCTCCGGCGGTCACCCCCAAAAGCAGGTCTCCAGCTTCTG | human VEGFA locus 3_VZ+434b_fwd_pKM600 |
| oIG9018 | CAGAAGCTGGACACGACTTCTTCAAGTCCGCCATGCCCGAGGGCTCCCATCACATC | +750 Target, Forward, biotinylated |
| oIG9019 | GATGTGATGGGAGCCCTCGGGCATGGCGGACTTGAAGAAGTCGTGTCCAGCTTCTG | +750 Target, Forward |
| oIG9024 | CAGAAGCTGGAAGAGCTCGTTTAGTGAACCGTCAGATCGCCGGGCTCCCATCACATC | CMVmin 1 Target, biotinylated |
| oIG9025 | GATGTGATGGGAGCCCGGCGATCTGACGGTTCACTAAACGAGCTCTTCCAGCTTCTG | CMVmin 1 Target |
| oIG9041 | CAGAAGCTGGAGTGTGCAGACGGCAGTCACTAGGGGGCGCTGGGCTCCCATCACATC | human VEGFA locus2, biotinylated |
| oIG9042 | GATGTGATGGGAGCCCAGCGCCCCCTAGTGACTGCCGTCTGCACACTCCAGCTTCTG | human VEGFA locus 2 |
| oIG9043 | CAGAAGCTGGACGTCTATATCACCGCCGACAAGCAGAAGAAGGGCTCCCATCACATC | target_nt950 pIG9003 fwd, biotinylated |
| oIG9044 | GATGTGATGGGAGCCCTTCTTCTGCTTGTCGGCGGTGATATAGACGTCCAGCTTCTG | target_nt950 pIG9003rev |
| | ||||
|---|---|---|---|---|
| plasmid name | Description | Backbone | Resistance | Reference |
| pIG0001 | dCas9, RFC 25 | pSB1C3 | Chloramph. | pX33a und pIG2004 |
| pIG0002 | pSV40, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0003 | SV40, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0004 | NLS, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0005 | Linker, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0006 | pSB1C3_VP16_pSB1C3 | pSB1C3 | cap | VP16 was PCR amplified from pMK018 (kindly provided by ...) using oIG0039 and oIG0040 and cloned into Xba1 and Spe1 pre cutted pSB1C3 vector using Gibson assambly. |
| pIG0007 | pSB1C3_KRAB_pSB1C3 | pSB1C3 | cap | VP16 was PCR amplified from pKM102 (kindly provided by ...) using oIG0041 and oIG0042 and cloned into Xba1 and Spe1 pre cutted pSB1C3 vector using Gibson assambly. |
| pIG0008 | G9a, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0009 | ERT2, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0010 | PIF, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0011 | PhyB, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0012 | UVR8, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0013 | COP1, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0014 | CIB, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0015 | CRY2, RFC 25 | pSB1C3 | Chloramph. | |
| pIG0016 | RNA Plasmid | pSB1C3 | Chloramph. | |
| pIG0017 | HA-Tag | pSB1C3 | Chloramph. | HA-Tag were designt as Oligos with overhangs to prefix and suffix of RFC25 (NgoMIV and SpeI sticky ends) |
| pIG0021 | pSB1C3_tracrRNA_crRNA_psB1C3 | pSB1C3 | Chloramph. | |
| pIG0018 | NLS | pSB1C3 | Chloramph. | NLS was used from FreiGEM10 (BBa_K404153, RFC25) |
| pIG0019 | CMV | pSB1C3 | Chloramph. | CMV was used from parts regestry (BBa_K747096, RFC10) |
| pIG0020 | BGH | pSB1C3 | Chloramph. | BGH was amplified from pGR2 with oIG0049 + oIG0050 and ligated into pSB1C3 via Gibson assembly. |
| pIG0021 a | NLS-dCas9 | pSB1C3 | Chloramph. | Ligation of pIG0018 and pIG0001 (RFC25) |
| pIG0022 | CMV-HA | pSB1C3 | Chloramph. | Ligation of pIG0017 and pIG0019 (RFC25) |
| pIG0023 | SV40-HA | pSB1C3 | Chloramph. | Ligation of pIG0003 and pIG0017 (RFC25) |
| pIG0024 | NLS-BGH | pSB1C3 | Chloramph. | Ligation of pIG0018 and pIG0020 (RFC25) |
| pIG0025 | CMV-HA-NLS-dCas9 | pSB1C3 | Chloramph. | Ligation of pIG0022 and pIG0021 (RFC25) |
| pIG0026 | SV40-HA-NLS-dCas9 | pSB1C3 | Chloramph. | Ligation of pIG0023 and pIG0021 (RFC25) |
| pIG0027 | CMV-HA-NLS-dCas9-Linker (3AS) | pSB1C3 | Chloramph. | Ligation of pIG0025 and pIG0029 (RFC25) |
| pIG0028 | CMV-HA-NLS-dCas9-Linker (7AS) | pSB1C3 | Chloramph. | Ligation of pIG0025 and pIG0030 (RFC25) |
| pIG0029 | SV40-HA-NLS-dCas9-Linker(3AS) | pSB1C3 | Chloramph. | Ligation of pIG0026 and pIG0029 (RFC25) |
| pIG0030 | SV40-HA-NLS-dCas9-Linker(7AS) | pSB1C3 | Chloramph. | Ligation of pIG0026 and pIG0030 (RFC25) |
| pIG0031 | Linker 3 AS | pSB1C3 | Chloramph. | Ligation of annealed oIG0045+46 and pSB1C3 RFC25 |
| pIG0032 | Linker (7 AS) | pSB1C3 | Chloramph. | Ligation of annealed oIG0051+52 and pSB1C3 RFC25 |
| pIG0033 | HA-NLS-dCas9-Linker (3AS) | pSB1C3 | Chloramph. | |
| pIG0034 | SV40 im 10er Standard | pSB1C3 | Chloramph. | |
| pIG0035 | BGH im 10er Standard | pSB1C3 | Chloramph. | |
| pIG0036 | HA-NLS-dCas9-Linker (3AS)-KRAB | pSB1C3 | Chloramph. | |
| pIG0037 | HA-NLS-dCas9-Linker (3AS)-G9a | pSB1C3 | Chloramph. | |
| pIG0038 | HA-NLS-dCas9-Linker (3AS)-VP16 | pSB1C3 | Chloramph. | |
| pIG0039 | HA-NLS-dCas9-Linker (3AS)-KRAB-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0040 | HA-NLS-dCas9-Linker (3AS)-G9a-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0041 | HA-NLS-dCas9-Linker (3AS)-VP16-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0042 | SV40-HA-NLS-dCas9-Linker (3AS)-KRAB-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0043 | SV40-HA-NLS-dCas9-Linker (3AS)-G9a-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0044 | SV40-HA-NLS-dCas9-Linker (3AS)-VP16-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0045 | CMV-HA-NLS-dCas9-Linker (3AS)-KRAB-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0046 | CMV-HA-NLS-dCas9-Linker (3AS)-G9a-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0047 | CMV-HA-NLS-dCas9-Linker (3AS)-VP16-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0048 | pSV40-PhyB | pSB1C3 | Chloramph. | |
| pIG0049 | pSV40-COP1 | pSB1C3 | Chloramph. | |
| pIG0050 | L3-VP16 | pSB1C3 | Chloramph. | |
| pIG0051 | L3-KRAB | pSB1C3 | Chloramph. | |
| pIG0052 | CMV-PhyB | pSB1C3 | Chloramph. | |
| pIG0053 | CMV-COP1 | pSB1C3 | Chloramph. | |
| pIG0054 | CMV-PhyB-L3-VP16 | pSB1C3 | Chloramph. | |
| pIG0055 | CMV-PhyB-L3-KRAB | pSB1C3 | Chloramph. | |
| pIG0056 | CMV-COP1-L3-VP16 | pSB1C3 | Chloramph. | |
| pIG0057 | CMV-COP1-L3-KRAB | pSB1C3 | Chloramph. | |
| pIG0058 | CMV-PhyB-L3-VP16-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0059 | CMV-PhyB-L3-KRAB-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0060 | CMV-COP1-L3-VP16-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0061 | CMV-COP1-L3-KRAB-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0062 | CMV-HA-NLS-dCas9-BGH | pSB1C3 | Chloramph. | |
| pIG0063 | SV40-HA-NLS-dCas9-BGH | pSB1C3 | Chloramph. | |
| pIG0064 | SV40-HA-NLS-dCas9-Linker 3AS-Uvr8 | pSB1C3 | Chloramph. | |
| pIG0065 | SV40-HA-NLS-dCas9-Linker 3AS-Piff | pSB1C3 | Chloramph. | |
| pIG0066 | SV40-HA-NLS-dCas9-Linker 3AS-Uvr8-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG0067 | SV40-HA-NLS-dCas9-Linker 3AS-Piff-NLS-BGH | pSB1C3 | Chloramph. | |
| pIG1000 | Siehe Liste unter Ornder "Aktivierung" | |||
| pIG1001 | Siehe Liste unter Ornder "Aktivierung" | |||
| pIG1002 | Siehe Liste unter Ornder "Aktivierung" | |||
| pIG1003 | Siehe Liste unter Ornder "Aktivierung" | |||
| pIG2000 | hU6 - precrRNA-site - CAG - dCas9, D10A | pX334a | Amp, Puro | Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong et al (Science. 2013 Jan 3. PubMed) |
| pIG2001 | UVR8 - KRAB - NLS | pKM102 | Amp, Tet | |
| pIG2002 | tetO13 - CMVmin - SEAP | pKM006 | Amp, ColE1 | Reporter Plasmid, in combination with pSAM200: constitutively high expression of SEAP |
| pIG2003 | tetR - VP16 | pSAM200 | Amp, ColE1 | Cis-activating fusion-protein of the used tetR-System |
| pIG2004 | hU6 - precrRNA-site - CAG - dCas9, D10A, H840A - KRAB | px334a | Amp, Puro | Zhang-plasmid, containing a H840A mutation, induced by side-directed mutagenesis (oIG2000 & 2001 / 2002 & 2003). Furtherly containing a KRAB domain, which originated from PCR amplification and subsequent Gibson Cloning of a pKM102 part (oIG2004 & oIG2005) with pX334a. |
| pIG2005 | hU6 - precrRNA-site - CAG - dCas9, D10A, H840A | pX334a | Amp, Puro | Zhang-plasmid, containing a H840A mutation, induced by side-directed mutagenesis (oIG2000-oIG2001, oIG2002-oIG2007). Gibson cloned of three PCR-fragments: oIG2000-oIG2001, oIG2002-oIG2007, oIG2008-oIG2009 |
| pIG2006 | hU6 - CMVmin-crRNA - HA - NLS - dCas9 - NLS | pX334 | Amp | - |
| pIG2007 | hU6 - CMV-crRNA - HA - NLS - dCas9 - NLS | pX334 | Amp | - |
| pIG2008 | hU6 - +230-crRNA - HA - NLS - dCas9 - NLS | pX334 | Amp | - |
| pIG2009 | hU6 - EMX-crRNA - HA - NLS - dCas9 - NLS | pX334 | Amp | - |
| pIG2010 | hU6 - CMVmin-crRNA - HA - NLS - dCas9 - KRAB - NLS | pX334 | Amp | - |
| pIG2011 | hU6 - CMV-crRNA - HA - NLS - dCas9 - KRAB - NLS | pX334 | Amp | - |
| pIG2012 | hU6 - +230-crRNA - HA - NLS - dCas9 - KRAB - NLS | pX334 | Amp | - |
| pIG2013 | hU6 - EMX-crRNA - HA - NLS - dCas9 - KRAB - NLS | pX334 | Amp | - |
| pIG2014 | hU6 - CMVmin-crRNA - HA - NLS - dCas9_H840A - NLS | pX334 | Amp | - |
| pIG2015 | hU6 - CMV-crRNA - HA - NLS - dCas9_H840A - NLS | pX334 | Amp | - |
| pIG2016 | hU6 - +230-crRNA - HA - NLS - dCas9_H840A - NLS | pX334 | Amp | - |
| pIG2017 | hU6 - EMX-crRNA - HA - NLS - dCas9_H840A - NLS | pX334 | Amp | - |
| pIG2018 | hU6 - +30-crRNA - HA - NLS - dCas9 - NLS | pX334 | Amp | - |
| pIG2019 | hU6 - +30-crRNA - HA - NLS - dCas9 - KRAB - NLS | pX334 | Amp | |
| pIG2020 | hU6 - +30-crRNA - HA - NLS - dCas9_H840A - NLS | pX334 | Amp | |
| pIG2021 | hU6 - Vegf II-crRNA - HA - NLS - dCas9 - KRAB - NLS | pX334 | Amp | |
| pIG2022 | hU6 - Vegf III-crRNA - HA - NLS - dCas9 - KRAB - NLS | pX334 | Amp | |
| pIG3001 | dCas9-mCherry | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333; Assembly was performed using Gibson PCR |
| pIG3002 | HA-NLS-dCas9-mCherry-NLS | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; HA-NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, the C-terminal NLS (different codon usage) was intoduced with a primer overlap; Assembly was performed using Gibson PCR |
| pIG3003 | HA-dCas9-mCherry-NLS | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, the C-terminal NLS (different codon usage) and HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR |
| pIG3004 | HA-NLS-dCas9-mCherry | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; HA-NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333; Assembly was performed using Gibson PCR |
| pIG3005 | HA-ERT2-NLS-dCas9-mCherry-NLS-ERT2 | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the C-terminal NLS (different codon usage) and the HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR |
| pIG3006 | HA-NLS-dCas9-mCherry-NLS-ERT2 | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; HA-NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the C-terminal NLS (different codon usage) was intoduced with a primer overlap; Assembly was performed using Gibson PCR |
| pIG3007 | HA-ERT2-NLS-dCas9-mCherry-NLS | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the C-terminal NLS (different codon usage) and the HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR |
| pIG3008 | HA-dCas9-mCherry-NLS-ERT2 | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the C-terminal NLS (different codon usage) and the HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR |
| pIG3009 | HA-ERT2-NLS-dCas9-mCherry | pMZ333 | Amp | Backbone was cut out of pMZ333 by NotI and XbaI; NLS-dCas9 was amplified by PCR (Primer with overlaps) from pX334a, mCherry from pMZ333, ERT2 from pMItom, the HA-Tag were intoduced with a primer overlap; Assembly was performed using Gibson PCR |
| pIG3010 | - | pX334a | amp | pX334a was digested with PstI to get rid of dCas9 and religated |
| pIG3011 | SEAP-Targetsequence I | pX334a | amp | pIG3010 was opened with BbsI and SEAP-Targetsequence I was inserted (oIG3013 + 14) |
| pIG3012 | SEAP-Targetsequence II | pX334a | amp | pIG3010 was opened with BbsI and SEAP-Targetsequence II was inserted (oIG3015 + 16) |
| pIG4000 | oMZ700PhyB (1-605) - mCherry-sv40pA | bla,ColE1 | mCherry was first PCR amplified from pMZ300 with oligos oMZ701/oMZ706, further amplified with oligos oMZ703/oMZ707, and Gibson-cloned into pMZ331 cut with XbaI/EcoRI | |
| pIG4001 | PhyB (1-908) - mCherry- sV40pA | bla,ColE1 | mCherry was first PCR amplified from pMZ300 with oligos oMZ701/oMZ706, further amplified using the oligos oMZ704/oMZ707 and Gibson-cloned into pMZ358 cut with XbaI/EcoRI. | |
| pIG4002 | hU6 - precrRNA-site - CAG - dCas9, D10A | AmpR | Multiplex Genome Engineering Using CRISPR/Cas Systems. Cong et al (Science. 2013 Jan 3. PubMed) | |
| pIG4003 | PIF3- mEGFP-sV40pA | bla,ColE1 | mEGFP was first PCR amplified from pMZ1050 with oligos oMZ728/oMZ729 and backbone pMZ354 was PCR amplified with oMZ726/oMZ727 for Gibson-Cloning. | |
| pIG4004 | PhyB(1-605)- mCherry- dCas9-sV40pA | pMZ700 | bla,ColE1 | mCherry was amplified from pMZ700 with oligos oHB058/ oHB059, dCas9 (hSpCsn1) was amplified from pMZ1025 with oHB060/ oHB061 and Gibson- Cloned into pMZ700 (after digestion of with XbaI/ EcorI |
| pIG4005 | PhyB(1-908)- mCherry- dCas9-sV40pA | pMZ701 | bla,ColE1 | mCherry was amplified from pIG4001 with oligos oIG4004/ oIG4001, dCas9 (hSpCsn1) was amplified from pHB1025 with oIG4002/ oIG4003 and Gibson- Cloned into pMZ701 (after digestion of with XbaI/ EcorI |
| pIG4006 | PhyB(1-908)- mCherry- dCas9 (with NLS)-sV40pA | pMZ701 | bla,ColE1 | mCherry was amplified from pIG4001 with oligos oIG4004/ oIG4001, dCas9 (hSpCsn1) was amplified from pIG4002 with oIG4002/ oIG4005 (+NLS!) and Gibson- Cloned into pIG4001(after digestion of pIG4001 with XbaI/ EcorI) |
| pIG4007 | PhyB(1-650)- mCherry- dCas9 (with NLS)-sV40pA | pMZ701 | bla,ColE1 | mCherry was amplified from pIG4000 with oligos oIG4000/ oIG4001, dCas9 (hSpCsn1) was amplified from pHB1025 with oIG4002/ oIG4005 and Gibson- Cloned into pIG4000 (after digestion of pIG4000 with XbaI/ EcorI |
| pIG6000 | pSb1C3_-Nhe1-pCMV-Sac2-eGFP-SalI-BanH1-NLS-terminator-Hind3-Kpn1-pSb1C3 | CMV promotor was amplified from Plasmid golgi-Gfp 405(kindly provided by toolbox) using oIG6000 and oIG6001. Avgfp was amplified from Plasmid golgi-Gfp 405 using oIG6002 and oIG6003. (iGEM cutting site was eliminated) BamH1 and Sal1were introduced to provide restrcitionsites for the insertion of caspase cleavage sites. a NLS was introduced. the terminator was amplified from Plasmid pmh 105 using oIG6022 and oIG6023. thereby Kpn1 und Nhe1 and the iGEM restrcitionsites Spe1 and Not1 were reestablished. Plasmid pSB1C3 serving as backbone was cut with Xba1 and Pst1. The 4 construts were fused together with a Gibson Assemby approach. | ||
| pIG6001 | pSb1C3_-Nhe1-pCMV-Sac2-CFP-SalI-BanH1-NLS-terminator-Hind3-Kpn1-pSb1C3 | CMV promotor was amplified from Plasmid golgi-Gfp 405(kindly provided by toolbox) using oIG6000 and oIG6001. ecfp was amplified from Plasmid pal 113 using oIG6002 and oIG6003. (iGEM cutting site was eliminated) BamH1 and Sal1were introduced to provide restrcitionsites for the insertion of caspase cleavage sites. a NLS was introduced. the terminator was amplified from Plasmid pmh 105 using oIG6022 and oIG6023. thereby Kpn1 und Nhe1 and the iGEM restrcitionsites Spe1 and Not1 were reestablished. Plasmid pSB1C3 serving as backbone was cut with Xba1 and Pst1. The 4 construts were fused together with a Gibson Assemby approach. | ||
| pIG8002 | HA-NLS-dCas9-GSAGSAG-G9a_NLS | pX334a | amp | |
| pIG8002_1 | pIG8002 cut with BbsI and ligated with VEGF crRNA1 | |||
| pIG8002_2 | pIG8002 cut with BbsI and ligated with VEGF crRNA2 | |||
| pIG8002_3 | pIG8002 cut with BbsI and ligated with VEGF crRNA3 | |||
| pIG8002_4 | pIG8002 cut with BbsI and ligated with VEGF crRNA4 | |||
| pIG8004 | HA-NLS-dCas9-GSAGSAG-dG9a_NLS | pX334a | amp | |
| pIG8004_1 | pIG8004 cut with BbsI and ligated with VEGF crRNA1 | |||
| pIG8004_2 | pIG8004 cut with BbsI and ligated with VEGF crRNA2 | |||
| pIG8004_3 | pIG8004 cut with BbsI and ligated with VEGF crRNA3 | |||
| pIG8004_4 | pIG8004 cut with BbsI and ligated with VEGF crRNA4 | |||
| pIG4100 | tetO13 - CMV Promotor - SEAP - SV40 | T405 | Amp | |
| pIG4101 | pSV40 - NLS - dCas9 - NLS - Linker - PIF - SV40 | Amp | ||
| pIG4102 | pSV40 - PhyB - KRAB - SV40 | Amp | ||
| pIG4103 | pSV40 - NLS - dCas9 - NLS - Linker - UVR8 - SV40 | Amp | ||
| pIG4104 | pSV40 - COP1 - KRAB - SV40 | Amp | ||
| pIG4105 | pSV40 - NLS - dCas9 - NLS - Linker - CRY2 - SV40 | Amp | ||
| pIG4106 | pSV40 - CIB - VP16 - SV40 | Amp | ||
| pIG4107 | pSV40 - CIB - KRAB - SV40 | Amp | ||
| pIG4108 | crRNA tetO13 1.CC | Amp | ||
| pIG4109 | crRNA tetO13 2.CC | Amp | ||
| pIG9000 | hU6 - EMX-crRNA - HA - NLS - dCas9 - NLS | px334a | Amp | |
