LOCUS       sequence-107487-        6448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  harbours marker genes URA3 and MET17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6448)
  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 6448)
  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: pUM
FEATURES             Location/Qualifiers
     source          1..6448
                     /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             complement(376..1710)
                     /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"
     CDS             2107..2910
                     /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(3098..3557)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3235..3256)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3447..3466
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3683..3705
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3697..3714
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3698..3714
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3721..3740
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3721..3739
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3748..3855
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(3867..3885)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3906..3922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3906..3922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3919..3941)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3930..3946
                     /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(3954..3960,3961..3978,3979..3984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3999..4020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4137..4154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4308..4896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4388..4407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5067..5858,5859..5927))
                     /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     5690..5709
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5928..6032)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(6062..6448,1..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 tgttggcggt cgagcttgtg agagaaagta ggtttataca taattttaca
      361 actcattacg cacactcatg gtttttggcc agcgaaaaca gtttcaaaag attgctggaa
      421 gtctgcaata atgtcatcaa taaattcgat accaacagag acacgaatta agtccttggt
      481 aacaccagat gccaactttt ctttgtcatt taattgtttg tgggtagtga agtatggagc
      541 aatgactaag gtcttggcat caccaacatt ggccaagtta gaggcaagct ttaaattgtc
      601 aacaacttga gcaccagaaa gtttgaatgg gtcagtttcc ttgtcggcat ttggtaagtc
      661 ttttacaccg aaagataaga caccaccgaa accgttagat agatacttct tagcattttc
      721 atgatgagaa tgagatgcta aaccagggta tgaaacccaa gatacgtatg gggattgttc
      781 taaccatttg gctaacttca atgcattttc accgtgtctt tcagctctca aagataatgt
      841 ttcaacacct tgtagtagca agaaagaggc aaatgggttc atcaatggac ccaaatctct
      901 taatagttca gttctaacat gaacgatgta tgccaagtta ccgtaggctt cattgtagat
      961 agtaccgtga tatccttcgg caggttgaga gaattgaggg aacttttctg ggtagtcctt
     1021 ccatgggaac ttaccagagt caacaataat accaccgata gtagtaccat gaccaccaat
     1081 ccatttggta gcagaatgtg ttacaatatc agcaccgtat ttaattggct gacagaagta
     1141 accaccggca ccaaatgtgt tgtcaacgac aactggaata ccgtgtttgt gagcaattgc
     1201 aacaattttt tcaaaatccg gaacattgta ctttggatta ccaatggttt ccaaataaac
     1261 agccttggtt ctttcatcaa agaccttttc gaattcttct ggattgtcac cttcaacaaa
     1321 tctagcctcg ataccaaatc ttttgaacga gattttgaac tggttataag taccaccgta
     1381 taagtaagaa gtggaaacga tgttgtcacc agtgtgtgcc aaaccttgga tggcaagggt
     1441 ttgagcggct tgaccggagg aaacagccaa agcagcagca ccaccttcta aagcagcaat
     1501 tctttcttcc aaaacattac tggttgggtt ttggaaacgg gaatagacgt aacctggaac
     1561 ttctagacca aacaattgcg aaccatgctt agagttttcg aaaacataag aagtggtggc
     1621 gtaaattggt acagctctgg atctgtgagc attgtcacca gggttctctt ggccggcgtg
     1681 tagttgaaca gtatcgaaat gagatggcat tgtatggatg ggggtaatag aattgtatct
     1741 atgtatctga cgaccctgta ttacacgaag gaaatagtag atgaaaagac aagagagcaa
     1801 gaaaaaggaa aaacttcctt tctaacagac gctttactta accttatata tatcttattt
     1861 ttttcatcga gcgtgttcaa ttggacaagg tgccaccttt tcgacacttc ggttattatg
     1921 ttacacagtt ttcatgagga tggcgcatgc gattcggtaa tctccgaaca gaaggaagaa
     1981 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     2041 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga
     2101 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     2161 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     2221 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     2281 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     2341 gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt
     2401 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     2461 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     2521 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     2581 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     2641 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     2701 attatgacac ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga
     2761 accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt
     2821 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     2881 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     2941 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga
     3001 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg gatcctatgc ggtgtgaaat
     3061 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     3121 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3181 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3241 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3301 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3361 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3421 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3481 ctggctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3541 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     3601 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     3661 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg
     3721 taatacgact cactataggg cgaattggag ctccaccgcg gtggcggccg ctctagaact
     3781 agtgggtccc ccgggctgca ggaattcgat atcaagctta tcgataccgt cgacctggag
     3841 ggggggcccg gtaccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
     3901 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3961 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4021 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4081 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4141 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4261 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4321 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4381 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4441 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4501 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4561 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4621 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4681 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4741 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4801 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4861 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4921 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4981 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5041 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5101 gcgatctgtc tatttcgttc atccatagtt gcctgactgc ccgtcgtgta gataactacg
     5161 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5221 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5281 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5341 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5401 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5461 tgatccccca tgttgtgaaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5521 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5581 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5641 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5701 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5761 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5821 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5881 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5941 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6001 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6061 ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa taatttggga
     6121 atttactctg tatttattta tttttatgtt ttgtatttgg attttagaaa gtaaataaag
     6181 aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac
     6241 aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa tagatataca
     6301 ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt cttctacaca
     6361 gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga taaaaggtag
     6421 tatttgttgg cgatccccct agagtctt
//
