LOCUS       sequence-107484-        6063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  harbours marker genes HIS3 and URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  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 6063)
  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: pHU
FEATURES             Location/Qualifiers
     source          1..6063
                     /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        509..696
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             697..1359
                     /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             1718..2521
                     /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(2713..3172)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2850..2871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3062..3081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3298..3320
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3312..3329
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3313..3329
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3336..3355
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3336..3354
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3363..3470
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(3482..3500)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3521..3537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3521..3537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3534..3556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3545..3561
                     /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(3569..3575,3576..3593,3594..3599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3614..3635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3752..3769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3923..4511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4003..4022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4682..5473,5474..5542))
                     /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     5305..5324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5543..5647)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(5677..6063,1..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 gcaccataaa ttcccgtttt aagagcttgg tgagcgctag gagtcactgc
      421 caggtatcgt ttgaacacgg cattagtcag ggaagtcata acacagtcct ttcccgcaat
      481 tttctttttc tattactctt ggcctcctct agtacactct atattttttt atgcctcggt
      541 aatgattttc attttttttt ttccacctag cggatgactc tttttttttc ttagcgattg
      601 gcattatcac ataatgaatt atacattata taaagtaatg tgatttcttc gaagaatata
      661 ctaaaaaatg agcaggcaag ataaacgaag gcaaagatga cagagcagaa agccctagta
      721 aagcgtatta caaatgaaac caagattcag attgcgatct ctttaaaggg tggtccccta
      781 gcgatagagc actcgatctt cccagaaaaa gaggcagaag cagtagcaga acaggccaca
      841 caatcgcaag tgattaacgt ccacacaggt atagggtttc tggaccatat gatacatgct
      901 ctggccaagc attccggctg gtcgctaatc gttgagtgca ttggtgactt acacatagac
      961 gaccatcaca ccactgaaga ctgcgggatt gctctcggtc aagcttttaa agaggcccta
     1021 ggggccgtgc gtggagtaaa aaggtttgga tcaggatttg cgcctttgga tgaggcactt
     1081 tccagagcgg tggtagatct ttcgaacagg ccgtacgcag ttgtcgaact tggtttgcaa
     1141 agggagaaag taggagatct ctcttgcgag atgatcccgc attttcttga aagctttgca
     1201 gaggctagca gaattaccct ccacgttgat tgtctgcgag gcaagaatga tcatcaccgt
     1261 agtgagagtg cgttcaaggc tcttgcggtt gccataagag aagccacctc gcccaatggt
     1321 accaacgatg ttccctccac caaaggtgtt cttatgtagt gacaccgatt atttaaagct
     1381 gcagcatacg atatatatac atgtgtatat atgtatacct atgaatgtca gtaagtatgt
     1441 atacgaacag tatgatactg aagatgacaa ggtaatgcat cattctatac gtgtcattct
     1501 gaacgaggcg cgctttcctt ttttcttttt gctttttctt tttttttctc ttgaactcga
     1561 cgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
     1621 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
     1681 gcacagaaca aaaacctgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
     1741 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
     1801 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
     1861 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
     1921 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
     1981 cttcgaagac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
     2041 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
     2101 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
     2161 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
     2221 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
     2281 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
     2341 caagggagac gcattgggtc aacagtatag aaccgtggat gatgtggtct ctacaggatc
     2401 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
     2461 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     2521 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     2581 attatatcag ttattaccct atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2641 accgcatcag gcgcggatcc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     2701 taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2761 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2821 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2881 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2941 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3001 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3061 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc tggcaagtgt agcggtcacg
     3121 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc
     3181 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3241 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3301 agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat
     3361 tggagctcca ccgcggtggc ggccgctcta gaactagtgg gtcccccggg ctgcaggaat
     3421 tcgatatcaa gcttatcgat accgtcgacc tggagggggg gcccggtacc agcttttgtt
     3481 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4741 tagttgcctg actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag
     5101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5641 ttccgcgcac atttccccga aaagtgccac ctgacggatc gcttgcctgt aacttacacg
     5701 cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtattt atttattttt
     5761 atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag
     5821 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt
     5881 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta
     5941 aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta
     6001 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt
     6061 ctt
//
