LOCUS Exported 7193 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION harbours marker genes HIS3 and LEU2. ACCESSION . VERSION . KEYWORDS pHL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7193) 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 7193) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7193 /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" promoter 1806..2213 /gene="S. cerevisiae LEU2" /label=LEU2 promoter CDS 2214..3308 /codon_start=1 /gene="S. cerevisiae LEU2" /product="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /label=LEU2 /note="yeast auxotrophic marker" /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG DLGGSNSTTEVGDAVAEEVKKILA" primer_bind complement(3980..4001) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4192..4211 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4428..4450 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4442..4459 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4443..4459 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 4466..4485 /label=T7 /note="T7 promoter, forward primer" promoter 4466..4484 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 4493..4600 /label=MCS /note="pBluescript multiple cloning site" promoter complement(4612..4630) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4651..4667) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4651..4667) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4664..4686) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4675..4691 /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(4699..4729) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4744..4765 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4882..4899) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5053..5641) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5133..5152) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5812..6672) /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 6435..6454 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6673..6777) /gene="bla" /label=AmpR promoter misc_feature 6807..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 gaactcgacg 1561 cgccatcgag gagaacttct agtatatcta catacctaat attattgcct tattaaaaat 1621 ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt cctgtacttc 1681 cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct atttctcaac 1741 aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat atcttgaccg 1801 cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc tcttagcaac 1861 cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac agaatcaaat 1921 tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata cctttttcaa 1981 ctgaaaaatt gggagaaaaa ggaaaggtga gagcgccgga accggctttt catatagaat 2041 agagaagcgt tcatgactaa atgcttgcat cacaatactt gaagttgaca atattattta 2101 aggacctatt gttttttcca ataggtggtt agcaatcgtc ttactttcta acttttctta 2161 ccttttacat ttcagcaata tatatatata tatttcaagg atataccatt ctaatgtctg 2221 cccctaagaa gatcgtcgtt ttgccaggtg accacgttgg tcaagaaatc acagccgaag 2281 ccattaaggt tcttaaagct atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa 2341 atcatttaat tggtggtgct gctatcgatg ctacaggtgt tccacttcca gatgaggcgc 2401 tggaagcctc caagaaggct gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg 2461 gtaccggtag tgttagacct gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt 2521 acgccaactt aagaccatgt aactttgcat ccgactctct tttagactta tctccaatca 2581 agccacaatt tgctaaaggt actgacttcg ttgttgtcag agaattagtg ggaggtattt 2641 actttggtaa gagaaaggaa gacgatggtg atggtgtcgc ttgggatagt gaacaataca 2701 ccgttccaga agtgcaaaga atcacaagaa tggccgcttt catggcccta caacatgagc 2761 caccattgcc tatttggtcc ttggataaag ctaatgtttt ggcctcttca agattatgga 2821 gaaaaactgt ggaggaaacc atcaagaacg aattccctac attgaaggtt caacatcaat 2881 tgattgattc tgccgccatg atcctagtta agaacccaac ccacctaaat ggtattataa 2941 tcaccagcaa catgtttggt gatatcatct ccgatgaagc ctccgttatc ccaggttcct 3001 tgggtttgtt gccatctgcg tccttggcct ctttgccaga caagaacacc gcatttggtt 3061 tgtacgaacc atgccacggt tctgctccag atttgccaaa gaataaggtc aaccctatcg 3121 ccactatctt gtctgctgca atgatgttga aattgtcatt gaacttgcct gaagaaggta 3181 aggccattga agatgcagtt aaaaaggttt tggatgcagg tatcagaact ggtgatttag 3241 gtggttccaa cagtaccacc gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc 3301 ttgcttaaaa agattctctt tttttatgat atttgtacat aaactttata aatgaaattc 3361 ataatagaaa cgacacgaaa ttacaaaatg gaatatgttc atagggtaga cgaaactata 3421 tacgcaatct acatacattt atcaagaagg agaaaaagga ggatgtaaag gaatacaggt 3481 aagcaaattg atactaatgg ctcaacgtga taaggaaaaa gaattgcact ttaacattaa 3541 tattgacaag gaggagggca ccacacaaaa agttaggtgt aacagaaaat catgaaacta 3601 tgattcctaa tttatatatt ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa 3661 aaacactcaa tgacctgacc atttgatgga gtttaagtca ataccttctt gaaccatttc 3721 ccataatggt gaaagttccc tcaagaattt tactctgtca gaaacggcct tacgacgtag 3781 tcgaggatcc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca 3841 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 3901 attttttaac caataggcca aaatcggcaa aatcccttat aaatcaaaag aatagaccga 3961 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 4021 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 4081 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 4141 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 4201 agcgaaagga gcgggcgcta gggcgctggc tggcaagtgt agcggtcacg ctgcgcgtaa 4261 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct 4321 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 4381 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 4441 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca 4501 ccgcggtggc ggccgctcta gaactagtgg gtcccccggg ctgcaggaat tcgatatcaa 4561 gcttatcgat accgtcgacc tggagggggg gcccggtacc agcttttgtt ccctttagtg 4621 agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4681 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4741 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4801 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 4861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 5041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5221 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5461 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 5521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 5641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 5701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 5761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 5821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 5881 actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 5941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 6001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 6061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 6121 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 6181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag cggttagctc 6241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 6301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 6361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 6421 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 6481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 6541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 6601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 6661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 6721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 6781 atttccccga aaagtgccac ctgacggatc gcttgcctgt aacttacacg cgcctcgtat 6841 cttttaatga tggaataatt tgggaattta ctctgtattt atttattttt atgttttgta 6901 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat 6961 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa 7021 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg 7081 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga 7141 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt ctt //