LOCUS       sequence-252828-        3846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 6 module- URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3846)
  AUTHORS   Goulev Y, Matifas A, Heyer V, Reina-San-Martin B, Charvin G
  TITLE     COSPLAY: An expandable toolbox for combinatorial and swift 
            generation of expression plasmids in yeast.
  JOURNAL   PLoS One. 2019 Aug 28;14(8):e0220694. doi: 
            10.1371/journal.pone.0220694. eCollection 2019.
  PUBMED    31461461
REFERENCE   2  (bases 1 to 3846)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2019-08-28"; volume: "14(8):e0220694. doi"; pages: " 
            10.1371/journal.pone.0220694. eCollection 2019"
COMMENT     Sequence Label: pMV10
FEATURES             Location/Qualifiers
     source          1..3846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..57
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        134..354
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             355..1158
                     /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(1296..1312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1296..1312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1309..1331)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1320..1336
                     /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(join(1344..1350,1351..1368,1369..1374))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1389..1410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1527..1544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1698..2286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1778..1797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2457..3248,3249..3317))
                     /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     3080..3099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3318..3422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3490..3508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3546..3568)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3668..3687
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
       61 accagtgaat tcgagctcgg tacctcgcga atgcatctag atatcggatc ccggtctcga
      121 gcaaccacag cttttcaatt caattcatca tttttttttt attctttttt ttgatttcgg
      181 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
      241 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
      301 gtattcttaa cccaactgca cagaacaaaa acctgcagga aacgaagata aatcatgtcg
      361 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
      421 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
      481 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
      541 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
      601 aagtacaatt ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa
      661 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
      721 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
      781 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
      841 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
      901 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat catgacaccc
      961 ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat
     1021 gtggtgtcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     1081 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     1141 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     1201 attagagctt caatttaatt atatcagtta ttaccctcca tagagaccgg gcccgtcgac
     1261 tgcagaggcc tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1321 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1381 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1441 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1501 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1561 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1621 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1681 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1741 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1801 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1861 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1921 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1981 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2041 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2101 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2161 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2221 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2281 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2341 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2401 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2461 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2521 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2581 gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag
     2641 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2701 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2761 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2821 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2881 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2941 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3001 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3061 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3121 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3181 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3241 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3301 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3361 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3421 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     3481 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     3541 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     3601 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     3661 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     3721 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     3781 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     3841 tgtgct
//
