LOCUS Exported 9068 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pCS4333 CEN/ARS vector, PGAL1-yeTadA1.0-100AA linker-T7 RNAP-TCYC1, TRP1 selectable marker. ACCESSION . VERSION . KEYWORDS pCS4333 yeTadA1.0-T7 RNAP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9068) TITLE Yeast expression plasmid for yeTadA1.0 - T7 RNAP fusion protein, enables targeted mutation downstream of T7 promoters JOURNAL Unpublished REFERENCE 2 (bases 1 to 9068) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9068 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(590..1450) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1213..1232 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1451..1555) /gene="bla" /label=AmpR promoter misc_feature 1592..2095 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" primer_bind 2137..2155 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2193..2215) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2315..2334 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 2351..2631 /gene="S. cerevisiae TRP1" /label=TRP1 promoter CDS 2632..3306 /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" rep_origin complement(3403..3858) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3540..3561) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3752..3771 /label=F1ori-R /note="F1 origin, reverse primer" terminator 3941..4106 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind 4027..4051 /label=pHybLex-R /note="ADH terminator, reverse primer" CDS complement(4254..4277) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter 4324..4988 /gene="GAL1 and GAL10 genes of S. cerevisiae" /label=GAL1,10 promoter /note="divergent inducible promoter, regulated by Gal4" protein_bind 4540..4657 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 4952..4975 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" terminator 8467..8656 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(8728..8745) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8899..419) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8979..8998) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 61 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 121 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 181 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 241 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 301 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 361 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 421 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 481 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 541 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 601 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 661 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 721 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 781 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 841 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 901 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 961 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 1021 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 1081 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 1141 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 1201 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 1261 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 1321 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 1381 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 1441 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 1501 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 1561 ttccccgaaa agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt 1621 aaatttttta atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa 1681 aaatagtttt tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac 1741 cttttatctt gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg 1801 tagaagacca cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta 1861 tatctattta atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg 1921 aaatttttta aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt 1981 atttactttc taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc 2041 aaattattcc atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtcc 2101 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt 2161 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg 2221 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg 2281 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga 2341 gtgcaccata aacgacatta ctatatatat aatataggaa gcatttaata gacagcatcg 2401 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat 2461 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga 2521 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga 2581 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt 2641 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag 2701 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat 2761 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat 2821 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag 2881 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag 2941 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca 3001 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg 3061 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg 3121 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca 3181 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt 3241 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag 3301 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg cctgcggtgt 3361 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata 3421 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg 3481 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc 3541 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa 3601 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt 3661 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac 3721 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta 3781 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg 3841 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc 3901 gatcggtgcg ggcctcttcg ctattacgcc agctgaattg gagcgacctc atgctatacc 3961 tgagaaagca acctgaccta caggaaagag ttactcaaga ataagaattt tcgttttaaa 4021 acctaagagt cactttaaaa tttgtataca cttatttttt ttataactta tttaataata 4081 aaaatcataa atcataagaa attcgcttat ttagaagtgt caacaacgta tctaccaacg 4141 atttgaccct tttccatctt ttcgtaaatt tctggcaagg tagacaagcc gacaaccttg 4201 attggagact tgaccaaacc tctggcgaag aattgttaat taagagctca gatcttatcg 4261 tcgtcatcct tgtaatccat cgatactagt gcggccgccc tttagtgagg gttgaattcg 4321 aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa taatcatatt 4381 acatggcatt accaccatat acatatccat atacatatcc atatctaatc ttacttatat 4441 gttgtggaaa tgtaaagagc cccattatct tagcctaaaa aaaccttctc tttggaactt 4501 tcagtaatac gcttaactgc tcattgctat attgaagtac ggattagaag ccgccgagcg 4561 ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt 4621 cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact 4681 agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat 4741 gaacgaatca aattaacaac cataggatga taatgcgatt agttttttag ccttatttct 4801 ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat aaatgcaaaa 4861 actgcataac cactttaact aatactttca acattttcgg tttgtattac ttcttattca 4921 aatgtaataa aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga 4981 gaaaaaaccc cggatccgat gtccgaggtt gagttcagcc atgagtactg gatgagacac 5041 gcattgactc tagcaaagag ggcgagagac gagagggaag tccccgtcgg cgccgtatta 5101 gtcttaaata atagggtaat tggtgaaggg tggaaccgtg ctattgggtt acatgacccg 5161 acagctcacg cggagattat ggcacttaga caaggcggtc ttgtaatgca gaactaccgt 5221 ttaatagacg caacactgta cgtcacattc gaaccttgtg taatgtgtgc cggagccatg 5281 atccactccc gtataggccg tgttgtcttt ggggtcagaa acgcgaagac aggagcagcc 5341 ggttccctga tggatgtgtt acactaccct ggaatgaatc atagtgtcga gataacggag 5401 ggtatcctgg cagacgagtg tgccgcgttg ctgtgttatt tttttaggat gccccgtcgg 5461 gtctttaatg cgcagaaaaa agcgcagccc tctaccggct ctggaggtgg aggttctgca 5521 gagtatgtgc gggccctctt tgactttaat gggaatgatg aagaagatct tccctttaag 5581 aaaggagaca tcctgagaat ccgggataag cctgaagagc agtggtggaa tgcagaggac 5641 agcgaaggaa agagggggat gattcctgtc ccttacgtgg agaagtattc cggagactat 5701 aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga tgacaatcag 5761 gttaacacga ttaacatagc taagaacgac ttctctgaca tcgaactggc tgctatcccg 5821 ttcaacactc tggctgacca ttacggtgag cgtttagctc gcgaacagtt ggcccttgag 5881 catgagtctt acgagatggg tgaagcacgc ttccgcaaga tgtttgagcg tcaacttaaa 5941 gctggtgagg ttgcggataa cgctgccgcc aagcctctca tcactaccct actccctaag 6001 atgattgcac gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg caagcgcccg 6061 acagccttcc agttcctgca agaaatcaag ccggaagccg tagcgtacat caccattaag 6121 accactctgg cttgcctaac cagtgctgac aatacaaccg ttcaggctgt agcaagcgca 6181 atcggtcggg ccattgagga cgaggctcgc ttcggtcgta tccgtgacct tgaagctaag 6241 cacttcaaga aaaacgttga ggaacaactc aacaagcgcg tagggcacgt ctacaagaaa 6301 gcatttatgc aagttgtcga ggctgacatg ctctctaagg gtctactcgg tggcgaggcg 6361 tggtcttcgt ggcataagga agactctatt catgtaggag tacgctgcat cgagatgctc 6421 attgagtcaa ccggaatggt tagcttacac cgccaaaatg ctggcgtagt aggtcaagac 6481 tctgagacta tcgaactcgc acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg 6541 ctggctggca tctctccgat gttccaacct tgcgtagttc ctcctaagcc gtggactggc 6601 attactggtg gtggctattg ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac 6661 agtaagaaag cactgatgcg ctacgaagac gtttacatgc ctgaggtgta caaagcgatt 6721 aacattgcgc aaaacaccgc atggaaaatc aacaagaaag tcctagcggt cgccaacgta 6781 atcaccaagt ggaagcattg tccggtcgag gacatccctg cgattgagcg tgaagaactc 6841 ccgatgaaac cggaagacat cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct 6901 gccgctgctg tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag ccttgagttc 6961 atgcttgagc aagccaataa gtttgctaac cataaggcca tctggttccc ttacaacatg 7021 gactggcgcg gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc 7081 aaaggactgc ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta ctactggctg 7141 aaaatccacg gtgcaaactg tgcgggtgtc gataaggttc cgttccctga gcgcatcaag 7201 ttcattgagg aaaaccacga gaacatcatg gcttgcgcta agtctccact ggagaacact 7261 tggtgggctg agcaagattc tccgttctgc ttccttgcgt tctgctttga gtacgctggg 7321 gtacagcacc acggcctgag ctataactgc tcccttccgc tggcgtttga cgggtcttgc 7381 tctggcatcc agcacttctc cgcgatgctc cgagatgagg taggtggtcg cgcggttaac 7441 ttgcttccta gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa agtcaacgag 7501 attctacaag cagacgcaat caatgggacc gataacgaag tagttaccgt gaccgatgag 7561 aacactggtg aaatctctga gaaagtcaag ctgggcacta aggcactggc tggtcaatgg 7621 ctggcttacg gtgttactcg cagtgtgact aagcgttcag tcatgacgct ggcttacggg 7681 tccaaagagt tcggcttccg tcaacaagtg ctggaagata ccattcagcc agctattgat 7741 tccggcaagg gtctgatgtt cactcagccg aatcaggctg ctggatacat ggctaagctg 7801 atttgggaat ctgtgagcgt gacggtggta gctgcggttg aagcaatgaa ctggcttaag 7861 tctgctgcta agctgctggc tgctgaggtc aaagataaga agactggaga gattcttcgc 7921 aagcgttgcg ctgtgcattg ggtaactcct gatggtttcc ctgtgtggca ggaatacaag 7981 aagcctattc agacgcgctt gaacctgatg ttcctcggtc agttccgctt acagcctacc 8041 attaacacca acaaagatag cgagattgat gcacacaaac aggagtctgg tatcgctcct 8101 aactttgtac acagccaaga cggtagccac cttcgtaaga ctgtagtgtg ggcacacgag 8161 aagtacggaa tcgaatcttt tgcactgatt cacgactcct tcggtaccat tccggctgac 8221 gctgcgaacc tgttcaaagc agtgcgcgaa actatggttg acacatatga gtcttgtgat 8281 gtactggctg atttctacga ccagttcgct gaccagttgc acgagtctca attggacaaa 8341 atgccagcac ttccggctaa aggtaacttg aacctccgtg acatcttaga gtcggacttc 8401 gcgttcgcgt aaaatcagta agcttggtac cgcggctagc taagcttggt accgcggcta 8461 gctaagatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat 8521 ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt 8581 ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt 8641 tttgggacgc tcgaagatcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 8701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 8761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 8821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 8881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 8941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 9001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 9061 ctttctcc //