LOCUS Exported 6063 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION harbours marker genes HIS3 and URA3. ACCESSION . VERSION . KEYWORDS pHU SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6063) 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 6063) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6063 /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 509..696 /gene="S. cerevisiae HIS3" /label=HIS3 promoter CDS 697..1359 /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 1718..2521 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" primer_bind complement(2850..2871) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3062..3081 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3298..3320 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3312..3329 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3313..3329 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3336..3355 /label=T7 /note="T7 promoter, forward primer" promoter 3336..3354 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 3363..3470 /label=MCS /note="pBluescript multiple cloning site" promoter complement(3482..3500) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3521..3537) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3521..3537) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3534..3556) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3545..3561 /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(3569..3599) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3614..3635 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3752..3769) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3923..4511) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4003..4022) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4682..5542) /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 5305..5324 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5543..5647) /gene="bla" /label=AmpR promoter misc_feature 5677..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 gcaccataaa ttcccgtttt aagagcttgg tgagcgctag gagtcactgc 421 caggtatcgt ttgaacacgg cattagtcag ggaagtcata acacagtcct ttcccgcaat 481 tttctttttc tattactctt ggcctcctct agtacactct atattttttt atgcctcggt 541 aatgattttc attttttttt ttccacctag cggatgactc tttttttttc ttagcgattg 601 gcattatcac ataatgaatt atacattata taaagtaatg tgatttcttc gaagaatata 661 ctaaaaaatg agcaggcaag ataaacgaag gcaaagatga cagagcagaa agccctagta 721 aagcgtatta caaatgaaac caagattcag attgcgatct ctttaaaggg tggtccccta 781 gcgatagagc actcgatctt cccagaaaaa gaggcagaag cagtagcaga acaggccaca 841 caatcgcaag tgattaacgt ccacacaggt atagggtttc tggaccatat gatacatgct 901 ctggccaagc attccggctg gtcgctaatc gttgagtgca ttggtgactt acacatagac 961 gaccatcaca ccactgaaga ctgcgggatt gctctcggtc aagcttttaa agaggcccta 1021 ggggccgtgc gtggagtaaa aaggtttgga tcaggatttg cgcctttgga tgaggcactt 1081 tccagagcgg tggtagatct ttcgaacagg ccgtacgcag ttgtcgaact tggtttgcaa 1141 agggagaaag taggagatct ctcttgcgag atgatcccgc attttcttga aagctttgca 1201 gaggctagca gaattaccct ccacgttgat tgtctgcgag gcaagaatga tcatcaccgt 1261 agtgagagtg cgttcaaggc tcttgcggtt gccataagag aagccacctc gcccaatggt 1321 accaacgatg ttccctccac caaaggtgtt cttatgtagt gacaccgatt atttaaagct 1381 gcagcatacg atatatatac atgtgtatat atgtatacct atgaatgtca gtaagtatgt 1441 atacgaacag tatgatactg aagatgacaa ggtaatgcat cattctatac gtgtcattct 1501 gaacgaggcg cgctttcctt ttttcttttt gctttttctt tttttttctc ttgaactcga 1561 cgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt 1621 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact 1681 gcacagaaca aaaacctgca ggaaacgaag ataaatcatg tcgaaagcta catataagga 1741 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa 1801 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt 1861 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt 1921 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact 1981 cttcgaagac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg 2041 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg 2101 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt 2161 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac 2221 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat 2281 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga 2341 caagggagac gcattgggtc aacagtatag aaccgtggat gatgtggtct ctacaggatc 2401 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg 2461 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta 2521 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta 2581 attatatcag ttattaccct atgcggtgtg aaataccgca cagatgcgta aggagaaaat 2641 accgcatcag gcgcggatcc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa 2701 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt 2761 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag 2821 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga 2881 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg 2941 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc 3001 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg 3061 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc tggcaagtgt agcggtcacg 3121 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc 3181 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc 3241 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc 3301 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat 3361 tggagctcca ccgcggtggc ggccgctcta gaactagtgg gtcccccggg ctgcaggaat 3421 tcgatatcaa gcttatcgat accgtcgacc tggagggggg gcccggtacc agcttttgtt 3481 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt 3541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 3601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 3721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 3781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 3841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 3901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 3961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 4081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 4141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 4201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 4261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 4321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 4381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 4441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 4501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 4561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 4621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 4681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 4741 tagttgcctg actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 4801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 4861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 4921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 4981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 5041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag 5101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 5161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 5221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 5281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 5341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 5401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 5461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 5521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 5581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 5641 ttccgcgcac atttccccga aaagtgccac ctgacggatc gcttgcctgt aacttacacg 5701 cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtattt atttattttt 5761 atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag 5821 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt 5881 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta 5941 aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta 6001 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt 6061 ctt //