LOCUS       sequence-107483-        7193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  harbours marker genes HIS3 and LEU2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7193)
  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 7193)
  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: pHL
FEATURES             Location/Qualifiers
     source          1..7193
                     /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"
     promoter        1806..2213
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2214..3308
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     rep_origin      complement(3843..4302)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3980..4001)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4192..4211
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4428..4450
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4442..4459
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4443..4459
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4466..4485
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4466..4484
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4493..4600
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4612..4630)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4651..4667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4651..4667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4664..4686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4675..4691
                     /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(4699..4705,4706..4723,4724..4729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4744..4765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4882..4899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5053..5641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5133..5152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5812..6603,6604..6672))
                     /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     6435..6454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6673..6777)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(6807..7193,1..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 gaactcgacg
     1561 cgccatcgag gagaacttct agtatatcta catacctaat attattgcct tattaaaaat
     1621 ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt cctgtacttc
     1681 cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct atttctcaac
     1741 aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat atcttgaccg
     1801 cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc tcttagcaac
     1861 cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac agaatcaaat
     1921 tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata cctttttcaa
     1981 ctgaaaaatt gggagaaaaa ggaaaggtga gagcgccgga accggctttt catatagaat
     2041 agagaagcgt tcatgactaa atgcttgcat cacaatactt gaagttgaca atattattta
     2101 aggacctatt gttttttcca ataggtggtt agcaatcgtc ttactttcta acttttctta
     2161 ccttttacat ttcagcaata tatatatata tatttcaagg atataccatt ctaatgtctg
     2221 cccctaagaa gatcgtcgtt ttgccaggtg accacgttgg tcaagaaatc acagccgaag
     2281 ccattaaggt tcttaaagct atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa
     2341 atcatttaat tggtggtgct gctatcgatg ctacaggtgt tccacttcca gatgaggcgc
     2401 tggaagcctc caagaaggct gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg
     2461 gtaccggtag tgttagacct gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt
     2521 acgccaactt aagaccatgt aactttgcat ccgactctct tttagactta tctccaatca
     2581 agccacaatt tgctaaaggt actgacttcg ttgttgtcag agaattagtg ggaggtattt
     2641 actttggtaa gagaaaggaa gacgatggtg atggtgtcgc ttgggatagt gaacaataca
     2701 ccgttccaga agtgcaaaga atcacaagaa tggccgcttt catggcccta caacatgagc
     2761 caccattgcc tatttggtcc ttggataaag ctaatgtttt ggcctcttca agattatgga
     2821 gaaaaactgt ggaggaaacc atcaagaacg aattccctac attgaaggtt caacatcaat
     2881 tgattgattc tgccgccatg atcctagtta agaacccaac ccacctaaat ggtattataa
     2941 tcaccagcaa catgtttggt gatatcatct ccgatgaagc ctccgttatc ccaggttcct
     3001 tgggtttgtt gccatctgcg tccttggcct ctttgccaga caagaacacc gcatttggtt
     3061 tgtacgaacc atgccacggt tctgctccag atttgccaaa gaataaggtc aaccctatcg
     3121 ccactatctt gtctgctgca atgatgttga aattgtcatt gaacttgcct gaagaaggta
     3181 aggccattga agatgcagtt aaaaaggttt tggatgcagg tatcagaact ggtgatttag
     3241 gtggttccaa cagtaccacc gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc
     3301 ttgcttaaaa agattctctt tttttatgat atttgtacat aaactttata aatgaaattc
     3361 ataatagaaa cgacacgaaa ttacaaaatg gaatatgttc atagggtaga cgaaactata
     3421 tacgcaatct acatacattt atcaagaagg agaaaaagga ggatgtaaag gaatacaggt
     3481 aagcaaattg atactaatgg ctcaacgtga taaggaaaaa gaattgcact ttaacattaa
     3541 tattgacaag gaggagggca ccacacaaaa agttaggtgt aacagaaaat catgaaacta
     3601 tgattcctaa tttatatatt ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa
     3661 aaacactcaa tgacctgacc atttgatgga gtttaagtca ataccttctt gaaccatttc
     3721 ccataatggt gaaagttccc tcaagaattt tactctgtca gaaacggcct tacgacgtag
     3781 tcgaggatcc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3841 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3901 attttttaac caataggcca aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3961 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     4021 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     4081 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     4141 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     4201 agcgaaagga gcgggcgcta gggcgctggc tggcaagtgt agcggtcacg ctgcgcgtaa
     4261 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
     4321 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     4381 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     4441 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
     4501 ccgcggtggc ggccgctcta gaactagtgg gtcccccggg ctgcaggaat tcgatatcaa
     4561 gcttatcgat accgtcgacc tggagggggg gcccggtacc agcttttgtt ccctttagtg
     4621 agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4681 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4741 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4801 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4861 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5221 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5461 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5701 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5761 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5821 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5881 actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5941 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6001 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6061 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6121 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6181 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag cggttagctc
     6241 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6301 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6361 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6421 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6481 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6541 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6601 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6661 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6781 atttccccga aaagtgccac ctgacggatc gcttgcctgt aacttacacg cgcctcgtat
     6841 cttttaatga tggaataatt tgggaattta ctctgtattt atttattttt atgttttgta
     6901 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat
     6961 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa
     7021 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg
     7081 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga
     7141 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt ctt
//
