LOCUS       sequence-407100-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Replicates in E. coli, S. cerevisiae, and A. laidlawii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  AUTHORS   Nucifora DP, Mehta ND, Giguere DJ, Karas BJ
  TITLE     An Expanded Genetic Toolbox to Accelerate the Creation of 
            Acholeplasma laidlawii Driven by Synthetic Genomes.
  JOURNAL   ACS Synth Biol. 2024 Jan 19;13(1):45-53. doi: 
            10.1021/acssynbio.3c00399. Epub 2023 Dec 19.
  PUBMED    38113213
REFERENCE   2  (bases 1 to 7083)
  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"; doi: 
            "10.1021/acssynbio.3c00399"; journalName: "ACS Synth Biol"; date: 
            "2024-01-19- 19"; volume: "13"; issue: "1"; pages: "45-53"
COMMENT     Sequence Label: pNZ18-CAH
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    56..559
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(4169..4180)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(6287..6949)
                     /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        complement(join(6950..7083,1..55))
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 aaaaaaaaaa aatgaaaatc attaccgagg cataaaaaaa tatagagtgt actaggatcg
       61 cttgcctgta acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac
      121 tctgtgttta tttattttta tgttttgtat ttggatttta gaaagtaaat aaagaaggta
      181 gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt caacaaaaag
      241 cgtactttac atatatattt attagacaag aaaagcagat taaatagata tacattcgat
      301 taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta cacagacaag
      361 atgaaacaat tcggcattaa tacctgagag caggaagagc aaggtaaaag gtagtatttg
      421 ttggcgatcc ccctagagtc ttttacatct tcggaaaaca aaaactattt tttctttaat
      481 ttcttttttt actttctatt tttaatttat atatttatat taaaaaattt aaattataat
      541 tatttttata gcacgtgatt gcagaaagag ctggtagttg gcgcactgtt cgaagaactg
      601 ccgatgtcca gtaagattct tactatgctg gttgaaccgg atgctggtaa agctacttgg
      661 gttgctgctt ctacttatgg taccgataca actactggtg aggaagttaa aggagctctt
      721 aaagaaatcc acttcagtag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
      781 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
      841 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
      901 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
      961 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     1021 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     1081 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     1141 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     1201 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     1261 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     1321 tgactgggtt gaaggctctc aagggcatcg gtcgacaagt tttgggattg ttaagggttc
     1381 cgaggctcaa cgtcaataaa gcaattggaa taaagaagcg aaaaaggaga agtcggttca
     1441 gaaaaagaag gatatggatc tggagctgta atataaaaac cttcttcaac taacggggca
     1501 ggttagtgac attagaaaac cgactgtaaa aagtacagtc ggcattatct catattataa
     1561 aagccagtca ttaggcctat ctgacaattc ctgaatagag ttcataaaca atcctgcatg
     1621 ataaccatca caaacagaat gatgtacctg taaagatagc ggtaaatata ttgaattacc
     1681 tttattaatg aattttcctg ctgtaataat gggtagaagg taattactat tattattgat
     1741 atttaagtta aacccagtaa atgaagtcca tggaataata gaaagagaaa aagcattttc
     1801 aggtataggt gttttgggaa acaatttccc cgaaccatta tatttctcta catcagaaag
     1861 gtataaatca taaaactctt tgaagtcatt ctttacagga gtccaaatac cagagaatgt
     1921 tttagataca ccatcaaaaa ttgtataaag tggctctaac ttatcccaat aacctaactc
     1981 tccgtcgcta ttgtaaccag ttctaaaagc tgtatttgag tttatcaccc ttgtcactaa
     2041 gaaaataaat gcagggtaaa atttatatcc ttcttgtttt atgtttcggt ataaaacact
     2101 aatatcaatt tctgtggtta tactaaaagt cgtttgttgg ttcaaataat gattaaatat
     2161 ctcttttctc ttccaattgt ctaaatcaat tttattaaag ttcatttgat atgcctccta
     2221 aatttttatc taaagtgaat ttaggaggct tacttgtctg ctttcttcat tagaatcaat
     2281 ccttttttaa aagtcaatat tactgtaaca taaatatata ttttaaaaat atcccacttt
     2341 atccaatttt cgtttgttga actaatgggt gctttagttg aagaataaaa gaccacatta
     2401 aaaaatgtgg tcttttgtgt ttttttaaag gatttgagcg tagcgaaaaa tccttttctt
     2461 tcttatcttg ataataaggg taactattgc cgggatagac tgtaacattc tcacgcataa
     2521 aatccccttt cattttctaa tgtaaatcta ttaccttatt attaattcaa ttcgctcata
     2581 attaatcctt tttcttatta cgcaaaatgg cccgatttaa gcacaccctt tattccgtta
     2641 atgcgccatg acagccatga taattactaa tactaggaga agttaataaa tacgtaacca
     2701 acatgattaa caattattag aggtcatcgt tcaaaatggt atgcgttttg acacatccac
     2761 tatatatccg tgtcgttctg tccactcctg aatcccattc cagaaattct ctagcgattc
     2821 cagaagtttc tcagagtcgg aaagttgacc agacattacg aactggcaca gatggtcata
     2881 acctgaagga agatctgatt gcttaactgc ttcagttaag accgaagcgc tcgtcgtata
     2941 acagatgcga tgatgcagac caatcaacat ggcacctgcc attgctacct gtacagtcaa
     3001 ggatggtaga aatgttgtcg gtccttgcac acgaatatta cgccatttgc ctgcatattc
     3061 aaacagctct tctacgataa gggcacaaat cgcatcgtgg aacgtttggg cttctaccga
     3121 tttagcagtt tgatacactt tctctaagta tccacctgaa tcataaatcg gcaaaataga
     3181 gaaaaattga ccatgtgtaa gcggccaatc tgattccacc tgagatgcat aatctagtag
     3241 aatctcttcg ctatcaaaat tcacttccac cttccactca ccggttgtcc attcatggct
     3301 gaactctgct tcctctgttg acatgacaca catcatctca atatccgaat agggcccatc
     3361 agtctgacga ccaagagagc cataaacacc aatagcctta acatcatccc catatttatc
     3421 caatattcgt tccttaattt catgaacaat cttcattctt tcttctctag tcattattat
     3481 tggtccattc actattctca ttcccttttc agataatttt agatttgctt ttctaaataa
     3541 gaatatttgg agagcaccgt tcttattcag ctattaataa ctcgtcttcc taagcatcct
     3601 tcaatccttt taataacaat tatagcatct aatcttcaac aaactggccc gtttgttgaa
     3661 ctactcttta ataaaataat ttttccgttc ccaattccac attgcaataa tagaaaatcc
     3721 atcttcatcg gctttttcgt catcatctgt atgaatcaaa tcgccttctt ctgtgtcatc
     3781 aaggtttaat tttttatgta tttcttttaa caaaccacca taggagatta accttttacg
     3841 gtgtaaacct tcctccaaat cagacaaacg tttcaaattc ttttcttcat catcggtcat
     3901 aaaatccgta tcctttacag gatattttgc agtttcgtca attgccgatt gtatatccga
     3961 tttatattta tttttcggtc gattttttat taaaacgtct caaaatcgtt tctgagacgt
     4021 tttagcgttt atttcgttta gttatcggca taatcgttaa aacaggcgtt atcgtagcgt
     4081 aaaagccctt gagcgtagcg tggctttgca gcgaagatgt tgtctgttag attatgaaag
     4141 ccgatgactg aatgaaataa taagcgcagc gtccttctat ttcggttgga ggaggctcaa
     4201 gggagtttga gggaatgaaa ttccctcatg ggtttgattt taaaaattgc ttgcaatttt
     4261 gccgagcggt agcgctggaa aatttttgaa aaaaatttgg aatttggaaa aaaatggggg
     4321 gaaaggaagc gaattttgct tccgtactac gaccccccat taagtgccga gtgccaattt
     4381 ttgtgccaaa aacgctctat cccaactggc tcaagggttt gaggggtttt tcaatcgcca
     4441 acgaatcgcc aacgttttcg ccaacgtttt ttataaatct atatttaagt agctttattg
     4501 ttgtttttat gattacaaag tgatacacta attttataaa attatttgat tggagttttt
     4561 taaatggtga tttcagaatc gaaaaaaaga gttatgattt ctctgacaaa agagcaagat
     4621 aaaaaattaa cagatatggc gaaacaaaaa ggtttttcaa aatctgcggt tgcggcgtta
     4681 gctatagaag aatatgcaag aaaggaatca gaacaaaaaa aataagcgaa agctcgcgtt
     4741 tttagaagga tacgagtttt cgctacttgt ttttgataag gtaatatatc atggctatta
     4801 aaaatactaa agctagaaat tttggatttt tattatatcc tgactcaatt cctaatgatt
     4861 ggaaagaaaa attagagagt ttgggcgtat ctatggctgt cagtccttta cacgatatgg
     4921 acgaaaaaaa agataaagat acatggaata gtagtgatgt tatacgaaat ggaaagcact
     4981 ataaaaaacc acactatcac gttatatata ttgcacgaaa tcctgtaaca atagaaagcg
     5041 ttaggaacaa gattaagcga aaattgggga atagttcagt tgctcatgtt gagatacttg
     5101 attatatcaa aggttcatat gaatatttga ctcatgaatc aaaggacgct attgctaaga
     5161 ataaacatat atacgacaaa aaagatattt tgaacattaa tgattttgat attgaccgct
     5221 atataacact tgatgaaagc caaaaaagag aattgaagaa tttactttta gatatagtgg
     5281 atgactataa tttggtaaat acaaaagatt taatggcttt tattcgcctt aggggagcgg
     5341 agtttggaat tttaaatacg aatgatgtaa aagatattgt ttcaacaaac tctagcgcct
     5401 ttagattatg gtttgagggc aattatcagt gtggatatag agcaagttat gcaaaggttc
     5461 ttgatgctga aacgggggaa ataaaatgac aaacaaagaa aaagagttat ttgctgaaaa
     5521 tgaggaatta aaaaaagaaa ttaaggactt aaaagagcgt attgaaagat acagagaaat
     5581 ggaagttgaa ttaagtacaa caatagattt attgagagga gggattattg aataaataaa
     5641 agcccccctg acgaaagtcg agctggatac ttcccgtccg ccagggggac atgccggcga
     5701 tgctgaaggt cgcgcgcatt cccgatgaag aggccggtta ccgcctgttt gaggatatag
     5761 taatctttct aaatagcttt ggattggagg agtatggcca ctaatactaa gtacaaacgt
     5821 gatgccattt ccataatgag ggatggtata aaatctaggt atagtaagga tggttgctgt
     5881 gctatatgtg gtagtagtga agacctagaa cttcatcact atcatactat atctcagcta
     5941 ataaaaaaat ttgctaaaga actccagctg gatttcactg atgagaatat cgtcggagat
     6001 aaatataata ttgttacgta cgctaatggc gtcaaaacaa agactctaga tggtggtatg
     6061 aaagctaaac cgtactccca aatatcctct tttaaagatc tgatcagctc cccgtttttc
     6121 acgcctccag ttaaatcccc ctttgtttta atccccatca aaattaaacg cgttagaatg
     6181 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     6241 ttctccggtg ccctgaatga actcaagtcc agactcctgt gtaaaactac ataagaacac
     6301 ctttggtgga gggaacatcg ttggtaccat tgggcgaggt ggcttctctt atggcaaccg
     6361 caagagcctt gaacgcactc tcactacggt gatgatcatt cttgcctcgc agacaatcaa
     6421 cgtggagggt aattctgcta gcctctgcaa agctttcaag aaaatgcggg atcatctcgc
     6481 aagagagatc tcctactttc tccctttgca aaccaagttc gacaactgcg tacggcctgt
     6541 tcgaaagatc taccaccgct ctggaaagtg cctcatccaa aggcgcaaat cctgatccaa
     6601 acctttttac tccacgcacg gcccctaggg cctctttaaa agcttgaccg agagcaatcc
     6661 cgcagtcttc agtggtgtga tggtcgtcta tgtgtaagtc accaatgcac tcaacgatta
     6721 gcgaccagcc ggaatgcttg gccagagcat gtatcatatg gtccagaaac cctatacctg
     6781 tgtggacgtt aatcacttgc gattgtgtgg cctgttctgc tactgcttct gcctcttttt
     6841 ctgggaagat cgagtgctct atcgctaggg gaccaccctt taaagagatc gcaatctgaa
     6901 tcttggtttc atttgtaata cgctttacta gggctttctg ctctgtcatc tttgccttcg
     6961 tttatcttgc ctgctcattt tttagtatat tcttcgaaga aatcacatta ctttatataa
     7021 tgtataattc attatgtgat aatgccaatc gctaagaaaa aaaaagagtc atccgctagg
     7081 tgg
//
