LOCUS       sequence-107486-        6588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  harbours marker genes HIS3 and MET17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6588)
  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 6588)
  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: pHM
FEATURES             Location/Qualifiers
     source          1..6588
                     /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"
     rep_origin      complement(3238..3697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3375..3396)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3587..3606
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3823..3845
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3837..3854
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3838..3854
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3861..3880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3861..3879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3888..3995
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4007..4025)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4046..4062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4046..4062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4059..4081)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4070..4086
                     /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(4094..4100,4101..4118,4119..4124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4139..4160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4277..4294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4448..5036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4528..4547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5207..5998,5999..6067))
                     /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     5830..5849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6068..6172)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(6202..6588,1..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 ctttcctttt 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 gatcctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     3241 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3301 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3361 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3421 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3481 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3541 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3601 aaggagcggg cgctagggcg ctggctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3661 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
     3721 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     3781 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     3841 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctccaccgcg
     3901 gtggcggccg ctctagaact agtgggtccc ccgggctgca ggaattcgat atcaagctta
     3961 tcgataccgt cgacctggag ggggggcccg gtaccagctt ttgttccctt tagtgagggt
     4021 taatttcgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4081 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4141 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4201 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4261 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4321 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4381 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4861 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactgc
     5281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5581 aacgatcaag gcgagttaca tgatccccca tgttgtgaaa aaaagcggtt agctccttcg
     5641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6181 cccgaaaagt gccacctgac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt
     6241 aatgatggaa taatttggga atttactctg tatttattta tttttatgtt ttgtatttgg
     6301 attttagaaa gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca
     6361 aaggtttaaa aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa
     6421 gcagattaaa tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt
     6481 cgtgtgtggt cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg
     6541 aagagcaaga taaaaggtag tatttgttgg cgatccccct agagtctt
//
