LOCUS       sequence-107495-        4822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  harbours marker gene URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4822)
  AUTHORS   Mulleder M, Campbell K, Matsarskaia O, Eckerstorfer F, Ralser M
  TITLE     Saccharomyces cerevisiae single-copy plasmids for auxotrophy 
            compensation, multiple marker selection, and for designing 
            metabolically cooperating communities.
  JOURNAL   F1000Res. 2016 Sep 20;5:2351. eCollection 2016.
  PUBMED    27830062
REFERENCE   2  (bases 1 to 4822)
  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: "F1000Res.";
            date: "2016-09-20"; volume: "5"; pages: "2351. eCollection 2016"
COMMENT     Sequence Label: pU
FEATURES             Location/Qualifiers
     source          1..4822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             477..1280
                     /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"
     rep_origin      complement(1472..1931)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1609..1630)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1821..1840
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2057..2079
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2071..2088
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2072..2088
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2095..2114
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2095..2113
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2122..2229
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(2241..2259)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2280..2296)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2280..2296)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2293..2315)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2304..2320
                     /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(2328..2334,2335..2352,2353..2358))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2373..2394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2511..2528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2682..3270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2762..2781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3441..4232,4233..4301))
                     /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     4064..4083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4302..4406)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(4436..4822,1..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 tgttggcggc gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      361 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      421 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      481 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      541 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      601 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      661 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      721 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      781 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      841 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      901 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      961 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     1021 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1081 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1141 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1201 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1261 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1321 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1381 agatgcgtaa ggagaaaata ccgcatcagg cgcggatcct atgcggtgtg aaataccgca
     1441 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa
     1501 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     1561 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     1621 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     1681 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     1741 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     1801 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggct
     1861 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     1921 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     1981 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     2041 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     2101 gactcactat agggcgaatt ggagctccac cgcggtggcg gccgctctag aactagtggg
     2161 tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct ggaggggggg
     2221 cccggtacca gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg
     2281 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2341 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2401 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2461 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2521 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2581 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2641 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2701 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2761 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2821 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2881 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2941 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3001 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3061 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3121 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3181 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3241 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3301 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3361 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3421 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3481 tgtctatttc gttcatccat agttgcctga ctgcccgtcg tgtagataac tacgatacgg
     3541 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3601 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3661 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3721 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3781 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3841 cccatgttgt gaaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3901 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3961 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4021 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4081 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4141 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4201 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4261 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4321 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4381 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacggatcg
     4441 cttgcctgta acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac
     4501 tctgtattta tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta
     4561 gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag
     4621 cgtactttac atatatattt attagacaag aaaagcagat taaatagata tacattcgat
     4681 taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag
     4741 atgaaacaat tcggcattaa tacctgagag caggaagagc aagataaaag gtagtatttg
     4801 ttggcgatcc ccctagagtc tt
//
