Team:TU Darmstadt/Materials/Primer Degradation

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Primer Degradation

Overview of the primer sequences used for the degradation project.

Name Sequnce (5' --> 3') Information
BBa EstA down GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGAAGTCCAGGCTCAGCGCCA Adds BioBrick compatible sites (up: EcoRI, NotI, XbaI / down: SpeI, NotI, PstI
BBa EstA up GTTTCTTCGAATTCGCGGCCGCTTCTAGATGGCTCCTTCGCCCTATTCGAC Adds BioBrick compatible sites (up: EcoRI, NotI, XbaI / down: SpeI, NotI, PstI
BBa FsC down GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAAGCAGAACCACGGACAGCCC Adds BioBrick compatible sites (up: EcoRI, NotI, XbaI / down: SpeI, NotI, PstI
BBa FsC up GTTTCTTCGAATTCGCGGCCGCTTCTAGATGCTGCCTACTTCTAACCCTGCC Adds BioBrick compatible sites (up: EcoRI, NotI, XbaI / down: SpeI, NotI, PstI
BBa PhoA down GTTTCTTCCTGCAGCGGCCGCTACTAGTAGTAGGCCGGCTGGGCCGCTTTTG Adds BioBrick compatible sites (up: EcoRI, NotI, XbaI / down: SpeI, NotI, PstI
BBa PhoA up GTTTCTTCGAATTCGCGGCCGCTTCTAGAGATGAAACAAAGCACTATTGCACTGGCAC Adds BioBrick compatible sites (up: EcoRI, NotI, XbaI / down: SpeI, NotI, PstI
BBa pNB-Est13 down GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAAATCTGCGGGTTCGGGGCTGTC Adds BioBrick compatible sites (up: EcoRI, NotI, XbaI / down: SpeI, NotI, PstI
BBa pNB-Est13 up GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTATGATACAACTGTCGAAACACGCTTC Adds BioBrick compatible sites (up: EcoRI, NotI, XbaI / down: SpeI, NotI, PstI
SKV a1 lo Ndel CGCGCGCATATGGGCCGGCTGGGCCGCTTTTG Adds restriction site to PhoA
SKV a1 up Xbal GCGCGCTCTAGAATGTTTACACTTTATGCTTCCG Adds restriction site to PhoA
SKV b1 lo Ncol CGCGCGCCATGGAATCTGCGGGTTCGGGGCTG Adds restriction site to Est13
SKV b1 up Ndel GCGCGCCATATGCATCACCATCACCACATGTATGATACAACTGTCGAAACACG Adds restriction site to Est13
SKV b2 lo Ncol CGCGCGCCATGGAGCAGAACCACGGACAGCCC Adds restriction site to FsC
SKV b2 up Ndel GCGCGCCATATGCATCACCATCACCACATGCTGCCTACTTCTAACCCTGC Adds restriction site to FsC
SKV c1 lo EcoRl CGCGCGGAATTCCTATCAGAAGTCCAGGCTCAGCGCCAGG Adds restriction site to EstA
SKV c1 up Ncol GCGCGCCCATGGGCTCCTTCGCCCTATTCGACGC Adds restriction site to EstA
SOE A up GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTTTACACTTTATGCTTCCGGCTCGTATG Adds BioBrick prefix to PhoA
SOE a1 lo TGTATCATACATGTGATGATGGTGGTGATGGGCCGGCTGGGCCGCTTTTG 20 bp overlapp PhoA His6 10 bp overlapp Est13
SOE a2 lo AGTAGGCAGCATGTGATGATGGTGGTGATGGGCCGGCTGGGCCGCTTTTG 20 bp overlapp PhoA His6 and 10 bp overlapp FsC
SOE b1 lo GTCGAATAGGGCGAAGGAGCGCCAATCTGCGGGTTCGGGG 20 bp overlapp EstA and Est13
SOE b1 up GCCCAGCCGGCCCATCACCACCATCATCACATGTATGATACAACTGTCGA 12 bp overlapp PhoA His6 und 20 bp overlapp Est13
SOE b2 lo GTCGAATAGGGCGAAGGAGCAGCAGAACCACGGACAGCCC 20 bp overlapp EstA and FsC
SOE b2 up GCCCAGCCGGCCCATCACCACCATCATCACATGCTGCCTACTTCTAACCC 12 bp overlapp PhoA His6 and 20 bp overlapp FsC
SOE c1 up CCCCGAACCCGCAGATTGGCGCTCCTTCGCCCTATTCGAC 20 bp overlapp Est13 and EstA
SOE c2 up GGGCTGTCCGTGGTTCTGCTGCTCCTTCGCCCTATTCGAC 20 bp overlapp FsC und EstA
SOE D lo GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGAAGTCCAGGCTCAGCGCC 20 overlapp EstA and adds BioBrick suffix
SOE Est13 mut lo CAATATCGAGAATCCTTAAGAA Mutates PstI site in Est13
SOE Est13 mut up CGATAAGCTTGCGAAAGCTGCACCAGAAAGATTCTTAAGGATTCTCGATA Mutates PstI site in Est13
SOE EstA PstI out up CGTGGTCTCCCTACAGCAGGCCGGCGCGCGCTAC Mutates PstI site in EstA
SOE EstA PstI out lo CCACGATGTAGCGCGCGCCGGCC Mutates PstI site in EstA
pEX FsC His SfiI up GCGCGCGGCCCAGCCGGCCCATCACCACCATCATCACATGCTGCCTACTTCTAACCC Adds SfiI site to ligate into a pEx expression system
pEX FsC Stop SfiI lo CGCGCGGGCCCCCGAGGCCGTTATCAAGCAGAACCACGGACAGCCCG Adds SfiI site to ligate into a pEx expression system
XbaI Rbs Phoa up GCGCGCTCTAGAGTTCACACAGGAAACCTACTAGATGAAACAAAGCACTATTGC Adds RBS to synthesis product: ENX-PhoA-His6tag-pNBEst13-Myctag-EstA-SNP
Est13 Bsa1 lo GCGCGCGGTCTCCAATCTGCGGGTTCGGGGCT Needed to amplify synthesis product ENX-PhoA-histag-pNBEst13-BsaI
EstA Bsa1 up GCGCGCGGTCTCAGATTGAACAAAAACTAATA Needed to ampflify snythesis product: BsaI-Myctag-EstA-SNP
EstA bba without stop codon tag lo TTTCTTCCTGCAGCGGCCGCTACTAGTATT Needed to ampflify snythesis product: BsaI-Myctag-EstA-SNP
M13 reverse up CAGGAAACAGCTATGAC Needed to get sequnce of the insert of the pEx-6His-tag plasmid
CalI pIII lo CCATTTTTCACTTCACAGGTCAAGC Needed to get sequnce of the insert of pEx-6His-tag plasmid