LOCUS       sequence-107480-        7687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  harbours marker genes HIS3, URA3 and MET17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7687)
  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 7687)
  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: pHUM
FEATURES             Location/Qualifiers
     source          1..7687
                     /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"
     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"
     CDS             complement(1611..2945)
                     /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             3342..4145
                     /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(4337..4796)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4474..4495)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4686..4705
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4922..4944
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4936..4953
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4937..4953
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4960..4979
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4960..4978
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4987..5094
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(5106..5124)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5145..5161)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5145..5161)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5158..5180)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5169..5185
                     /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(5193..5199,5200..5217,5218..5223))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5238..5259
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5376..5393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5547..6135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5627..5646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6306..7097,7098..7166))
                     /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     6929..6948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7167..7271)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(7301..7687,1..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 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 cttccctttt ttctttttgc tttttctttt tttttctctt gaactcgagc
     1561 ttgtgagaga aagtaggttt atacataatt ttacaactca ttacgcacac tcatggtttt
     1621 tggccagcga aaacagtttc aaaagattgc tggaagtctg caataatgtc atcaataaat
     1681 tcgataccaa cagagacacg aattaagtcc ttggtaacac cagatgccaa cttttctttg
     1741 tcatttaatt gtttgtgggt agtgaagtat ggagcaatga ctaaggtctt ggcatcacca
     1801 acattggcca agttagaggc aagctttaaa ttgtcaacaa cttgagcacc agaaagtttg
     1861 aatgggtcag tttccttgtc ggcatttggt aagtctttta caccgaaaga taagacacca
     1921 ccgaaaccgt tagatagata cttcttagca ttttcatgat gagaatgaga tgctaaacca
     1981 gggtatgaaa cccaagatac gtatggggat tgttctaacc atttggctaa cttcaatgca
     2041 ttttcaccgt gtctttcagc tctcaaagat aatgtttcaa caccttgtag tagcaagaaa
     2101 gaggcaaatg ggttcatcaa tggacccaaa tctcttaata gttcagttct aacatgaacg
     2161 atgtatgcca agttaccgta ggcttcattg tagatagtac cgtgatatcc ttcggcaggt
     2221 tgagagaatt gagggaactt ttctgggtag tccttccatg ggaacttacc agagtcaaca
     2281 ataataccac cgatagtagt accatgacca ccaatccatt tggtagcaga atgtgttaca
     2341 atatcagcac cgtatttaat tggctgacag aagtaaccac cggcaccaaa tgtgttgtca
     2401 acgacaactg gaataccgtg tttgtgagca attgcaacaa ttttttcaaa atccggaaca
     2461 ttgtactttg gattaccaat ggtttccaaa taaacagcct tggttctttc atcaaagacc
     2521 ttttcgaatt cttctggatt gtcaccttca acaaatctag cctcgatacc aaatcttttg
     2581 aacgagattt tgaactggtt ataagtacca ccgtataagt aagaagtgga aacgatgttg
     2641 tcaccagtgt gtgccaaacc ttggatggca agggtttgag cggcttgacc ggaggaaaca
     2701 gccaaagcag cagcaccacc ttctaaagca gcaattcttt cttccaaaac attactggtt
     2761 gggttttgga aacgggaata gacgtaacct ggaacttcta gaccaaacaa ttgcgaacca
     2821 tgcttagagt tttcgaaaac ataagaagtg gtggcgtaaa ttggtacagc tctggatctg
     2881 tgagcattgt caccagggtt ctcttggccg gcgtgtagtt gaacagtatc gaaatgagat
     2941 ggcattgtat ggatgggggt aatagaattg tatctatgta tctgacgacc ctgtattaca
     3001 cgaaggaaat agtagatgaa aagacaagag agcaagaaaa aggaaaaact tcctttctaa
     3061 cagacgcttt acttaacctt atatatatct tatttttttc atcgagcgtg ttcaattgga
     3121 caaggtgcca ccttttcgac acttcggtta ttatgttaca cagttttcat gaggatggcg
     3181 catgcgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     3241 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     3301 aactgcacag aacaaaaacc tgcaggaaac gaagataaat catgtcgaaa gctacatata
     3361 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     3421 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     3481 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     3541 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     3601 tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     3661 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     3721 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     3781 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     3841 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     3901 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     3961 atgacaaggg agacgcattg ggtcaacagt atagaaccgt ggatgatgtg gtctctacag
     4021 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     4081 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     4141 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     4201 tttaattata tcagttatta ccctatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4261 aaataccgca tcaggcgcgg atcctatgcg gtgtgaaata ccgcacagat gcgtaaggag
     4321 aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4381 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4441 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4501 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4561 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4621 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4681 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggctggcaa gtgtagcggt
     4741 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     4801 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     4861 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     4921 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     4981 gaattggagc tccaccgcgg tggcggccgc tctagaacta gtgggtcccc cgggctgcag
     5041 gaattcgata tcaagcttat cgataccgtc gacctggagg gggggcccgg taccagcttt
     5101 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
     5161 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5221 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5281 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5341 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5401 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5461 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5521 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5581 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5641 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5701 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5761 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5821 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5881 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5941 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6001 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6061 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6121 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6181 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6241 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6301 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6361 tccatagttg cctgactgcc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6421 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6481 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6541 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6601 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6661 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgaaaa
     6721 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6781 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6841 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6901 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6961 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7021 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7081 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7141 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7201 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7261 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg gatcgcttgc ctgtaactta
     7321 cacgcgcctc gtatctttta atgatggaat aatttgggaa tttactctgt atttatttat
     7381 ttttatgttt tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat
     7441 gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata
     7501 tatttattag acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa
     7561 tgtaaaatca caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc
     7621 attaatacct gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta
     7681 gagtctt
//
