LOCUS Exported 6588 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION harbours marker genes HIS3 and MET17. ACCESSION . VERSION . KEYWORDS pHM SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6588) AUTHORS Michael Mulleder, Kate Campbell, Olga Matsarskaia, Florian Eckerstorfer, Markus Ralser TITLE Saccharomyces cerevisiae single-copy plasmids for auxotrophy compensation, multiple marker selection, and for designing metabolically cooperating communities JOURNAL F1000Research REFERENCE 2 (bases 1 to 6588) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6588 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 165..183 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(221..243) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 343..362 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 507..694 /gene="S. cerevisiae HIS3" /label=HIS3 promoter CDS 695..1357 /codon_start=1 /gene="S. cerevisiae HIS3" /product="imidazoleglycerol-phosphate dehydratase, required for histidine biosynthesis" /label=HIS3 /note="yeast auxotrophic marker" /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM" CDS complement(1611..2945) /codon_start=1 /gene="S. cerevisiae MET17" /gene_synonym="MET15; MET25" /product="O-acetylserine and O-acetylhomoserine sulfhydrylase, required for sulfur amino acid synthesis" /label=MET17 /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP" primer_bind complement(3375..3396) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3587..3606 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3823..3845 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3837..3854 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3838..3854 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3861..3880 /label=T7 /note="T7 promoter, forward primer" promoter 3861..3879 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 3888..3995 /label=MCS /note="pBluescript multiple cloning site" promoter complement(4007..4025) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4046..4062) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4046..4062) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4059..4081) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4070..4086 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4094..4124) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4139..4160 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4277..4294) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4448..5036) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4528..4547) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5207..6067) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5830..5849 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6068..6172) /gene="bla" /label=AmpR promoter misc_feature 6202..117 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" ORIGIN 1 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt 61 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa 121 aaggacccta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga 181 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc 241 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg 301 cgtcagcggg tgttggcgga cgtcggggct ggcttaacta tgcggcatca gagcagattg 361 tactgagagt gcaccataat tccgttttaa gagcttggtg agcgctagga gtcactgcca 421 ggtatcgttt gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt 481 tctttttcta ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa 541 tgattttcat tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc 601 attatcacat aatgaattat acattatata aagtaatgtg atttcttcga agaatatact 661 aaaaaatgag caggcaagat aaacgaaggc aaagatgaca gagcagaaag ccctagtaaa 721 gcgtattaca aatgaaacca agattcagat tgcgatctct ttaaagggtg gtcccctagc 781 gatagagcac tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac aggccacaca 841 atcgcaagtg attaacgtcc acacaggtat agggtttctg gaccatatga tacatgctct 901 ggccaagcat tccggctggt cgctaatcgt tgagtgcatt ggtgacttac acatagacga 961 ccatcacacc actgaagact gcgggattgc tctcggtcaa gcttttaaag aggccctagg 1021 ggccgtgcgt ggagtaaaaa ggtttggatc aggatttgcg cctttggatg aggcactttc 1081 cagagcggtg gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag 1141 ggagaaagta ggagatctct cttgcgagat gatcccgcat tttcttgaaa gctttgcaga 1201 ggctagcaga attaccctcc acgttgattg tctgcgaggc aagaatgatc atcaccgtag 1261 tgagagtgcg ttcaaggctc ttgcggttgc cataagagaa gccacctcgc ccaatggtac 1321 caacgatgtt ccctccacca aaggtgttct tatgtagtga caccgattat ttaaagctgc 1381 agcatacgat atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat 1441 acgaacagta tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga 1501 acgaggcgcg ctttcctttt ttctttttgc tttttctttt tttttctctt gaactcgagc 1561 ttgtgagaga aagtaggttt atacataatt ttacaactca ttacgcacac tcatggtttt 1621 tggccagcga aaacagtttc aaaagattgc tggaagtctg caataatgtc atcaataaat 1681 tcgataccaa cagagacacg aattaagtcc ttggtaacac cagatgccaa cttttctttg 1741 tcatttaatt gtttgtgggt agtgaagtat ggagcaatga ctaaggtctt ggcatcacca 1801 acattggcca agttagaggc aagctttaaa ttgtcaacaa cttgagcacc agaaagtttg 1861 aatgggtcag tttccttgtc ggcatttggt aagtctttta caccgaaaga taagacacca 1921 ccgaaaccgt tagatagata cttcttagca ttttcatgat gagaatgaga tgctaaacca 1981 gggtatgaaa cccaagatac gtatggggat tgttctaacc atttggctaa cttcaatgca 2041 ttttcaccgt gtctttcagc tctcaaagat aatgtttcaa caccttgtag tagcaagaaa 2101 gaggcaaatg ggttcatcaa tggacccaaa tctcttaata gttcagttct aacatgaacg 2161 atgtatgcca agttaccgta ggcttcattg tagatagtac cgtgatatcc ttcggcaggt 2221 tgagagaatt gagggaactt ttctgggtag tccttccatg ggaacttacc agagtcaaca 2281 ataataccac cgatagtagt accatgacca ccaatccatt tggtagcaga atgtgttaca 2341 atatcagcac cgtatttaat tggctgacag aagtaaccac cggcaccaaa tgtgttgtca 2401 acgacaactg gaataccgtg tttgtgagca attgcaacaa ttttttcaaa atccggaaca 2461 ttgtactttg gattaccaat ggtttccaaa taaacagcct tggttctttc atcaaagacc 2521 ttttcgaatt cttctggatt gtcaccttca acaaatctag cctcgatacc aaatcttttg 2581 aacgagattt tgaactggtt ataagtacca ccgtataagt aagaagtgga aacgatgttg 2641 tcaccagtgt gtgccaaacc ttggatggca agggtttgag cggcttgacc ggaggaaaca 2701 gccaaagcag cagcaccacc ttctaaagca gcaattcttt cttccaaaac attactggtt 2761 gggttttgga aacgggaata gacgtaacct ggaacttcta gaccaaacaa ttgcgaacca 2821 tgcttagagt tttcgaaaac ataagaagtg gtggcgtaaa ttggtacagc tctggatctg 2881 tgagcattgt caccagggtt ctcttggccg gcgtgtagtt gaacagtatc gaaatgagat 2941 ggcattgtat ggatgggggt aatagaattg tatctatgta tctgacgacc ctgtattaca 3001 cgaaggaaat agtagatgaa aagacaagag agcaagaaaa aggaaaaact tcctttctaa 3061 cagacgcttt acttaacctt atatatatct tatttttttc atcgagcgtg ttcaattgga 3121 caaggtgcca ccttttcgac acttcggtta ttatgttaca cagttttcat gaggatggcg 3181 gatcctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa 3241 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 3301 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag 3361 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 3421 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat 3481 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc 3541 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga 3601 aaggagcggg cgctagggcg ctggctggca agtgtagcgg tcacgctgcg cgtaaccacc 3661 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca 3721 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg 3781 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta 3841 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctccaccgcg 3901 gtggcggccg ctctagaact agtgggtccc ccgggctgca ggaattcgat atcaagctta 3961 tcgataccgt cgacctggag ggggggcccg gtaccagctt ttgttccctt tagtgagggt 4021 taatttcgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 4081 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 4141 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 4201 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 4261 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 4321 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 4381 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 4441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 4501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 4561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 4621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 4681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 4741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 4801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 4861 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 4921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 4981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 5041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 5101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 5161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactgc 5281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 5461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 5521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 5581 aacgatcaag gcgagttaca tgatccccca tgttgtgaaa aaaagcggtt agctccttcg 5641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 5701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 5761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 5821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 5881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 5941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 6001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 6061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 6121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6181 cccgaaaagt gccacctgac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt 6241 aatgatggaa taatttggga atttactctg tatttattta tttttatgtt ttgtatttgg 6301 attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca 6361 aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa 6421 gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt 6481 cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg 6541 aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctt //