LOCUS       sequence-254729-        7602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A nuclear plasmid encoding the WT TP-DNAP1 for OrthoRep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  AUTHORS   Ravikumar A, Arzumanyan GA, Obadi MKA, Javanpour AA, Liu CC
  TITLE     Scalable, Continuous Evolution of Genes at Mutation Rates above 
            Genomic Error Thresholds.
  JOURNAL   Cell. 2018 Dec 13;175(7):1946-1957.e13. doi: 
            10.1016/j.cell.2018.10.021. Epub 2018 Nov 8.
  PUBMED    30415839
REFERENCE   2  (bases 1 to 7602)
  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: 
            "10.1016/j.cell.2018.10.021"; journalName: "Cell"; date: 
            "2018-12-13- 13"; volume: "175"; issue: "7"; pages: "1946-1957.e13"
COMMENT     Sequence Label: pAR-Ec318
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    283..786
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     827..845
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(883..905)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1005..1024
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1217..1403
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1404..2066
                     /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"
     terminator      6011..6176
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6066..6090)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      complement(6219..6807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6299..6318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7021..7040)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(7032..7602,1..245)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(7032..7602,1..245)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(7105..7124)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
       61 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
      121 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
      181 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
      241 tctaatcaga attggttaat tggttgtaac acggtccttt tcatcacgtg ctataaaaat
      301 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
      361 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
      421 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
      481 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
      541 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
      601 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
      661 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
      721 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
      781 gcgatccgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
      841 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
      901 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
      961 cgcgtcagtg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     1021 tgtactgaga gtgcaccata ccaccttttc aattcatcat ttttttttta ttcttttttt
     1081 tgattttaat tccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
     1141 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
     1201 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
     1261 tttttttttc cacctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
     1321 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
     1381 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
     1441 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
     1501 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
     1561 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
     1621 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
     1681 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctaggg gccgtgcgtg
     1741 gagtaaaaag gtttggatca ggatttgcgc ctttggatga ggcactttcc agagcggtgg
     1801 ttgatctttc gaacaggccg tacgcagttg tcgaacttgg tttgcaaagg gagaaagtag
     1861 gtgatctctc ttgcgagatg atcccgcatt ttcttgaaag ctttgcagag gctagcagaa
     1921 ttaccctcca cgttgattgt ctgcgaggca agaatgatca tcaccgtagt gagagtgcgt
     1981 tcaaggctct tgcggttgcc ataagagaag ccacctcgcc caatggtacc aacgatgttc
     2041 cctccaccaa aggtgttctt atgtagtgac accgattatt taaagctgct gcatacgata
     2101 tatatacatg tgtatatatg tatacctatg aatgtcagta agtatgtata cgaacagtat
     2161 gatactgaag atgacaaggt aatgcatcat tctatacgtg tcattctgaa cgaggcgcgc
     2221 tttccttttt tctttttgct ttttcttttt ttttctcttg aactcgacgg atcatatatg
     2281 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggga gctcgtgttg
     2341 ttatccgata caaccggata tttttctttt aatgagtcta aaccgtgata gcttcaggtt
     2401 aatacaatca aaaaaagctc aaatattctt ttaatgccgc gttcacagat tccaattgaa
     2461 tacaactagg tagttcatta tatgaagcct ttgctactat ttttcactat agtctgcctt
     2521 caccttaatg cagacatcca catattttaa tcactttaaa ataaaaagga agatatatta
     2581 gaagctatga tccaatctgt aagccagatt aaaattcacg aactcttctt tcatttgaat
     2641 tgaatgcttt gagttggggt agattatcgc aaattactca tcacatttat tgactacgaa
     2701 cttgctgatg tccttttttt atttatattt ttcttcagtg aagcgatttt ttttttacac
     2761 agaccaagac ggaaaaaagt agctaaggaa gaaaacaaaa tcatgaaaaa aatgtgaagt
     2821 gatcatgcac atcgcatcaa cttaaacatt ggcttagaga tatatagagt tagagtttac
     2881 ggcaaccttt aagcaccaat accttttggc atagtctaaa gacctggttc ttaattttaa
     2941 acaaatttaa ctaaagattt ccctatcaaa gaagtaacga gttgacagat tttctcaaaa
     3001 taaatcgata ctgcatttct aggcatatcc agcgagatct atggactaca aagacaaagc
     3061 attaaacgac ttgagaaacg tatacgcaga cttcgactcc ttgccattgg acttcagaca
     3121 aatcttaatt aaggatagag ctacattgtt gcaaaaggaa gacgttgaaa agaaaatttt
     3181 ggaaagacaa gaagatgcca aaaagtacgc tgaatacttg aagcaatcag aaatcccaga
     3241 aagaatctcc ttacctaaca tcaagagaca taagggtgta tcaatctcct ttgaagaaac
     3301 atctgaagat atggtcttgg aaccaagacc ttttatcttc gacggtttga acatcagatg
     3361 tttcagaaga gaaaccatat tcagtttgaa aaataagatt ttgaacatgg ttaaggaatc
     3421 ttcatccttt aaaaacgttt caagacaaag tgtatctttt atgtacttca aaatttttaa
     3481 taagggtaaa gttattgcaa gtacaaagtc tgttaacatc tatgaagata agatcgacga
     3541 aagattggaa gatttgtgta acaacttcga tgacgtattg aaaaagatta tagacgtcac
     3601 ctatggttac gaatccttat tcgttagtga aacttactct tacgtaatat tctatgctaa
     3661 atcaatctac tttccacaac ctagatgcgt taacaactgg ggtaacaaca tcccaaacat
     3721 cttgacattc gattccttta agcttttcac cgcaaacaaa aacaacgtta gttgtatcaa
     3781 gcaatgttca agatttttgt ggcaaaaaga tttcaacact ttagaagaaa tgatagaata
     3841 caagaacggt aatatttgta tcgtcacacc acaattgcat atcaacgatg ttagagacat
     3901 caagtcattc aacgatatca gattgtactc agaatcccct attaaaacat tttccgttat
     3961 tgataacact atcacatatt tgttttactt caaggaacat ttgggtgtta tttttaacat
     4021 cactaagtca agacatgata gaagagtaac aaaattttca ccattgtcca agttcagtga
     4081 cgtcaaaaat atcactgttt gtttcgatat cgaaagttac ttcgacccag aaaaggaatc
     4141 taaccaagtt aacatccctt tcatttgttg cgcttcaatc atctataata aggttattgg
     4201 taacatcgtc gattttgaag gtagagactg tgtcgcacaa atgatcgaat acgttgtaga
     4261 tatttgcggt gaattgaaca taagttctgt tgaattgatc gcccataacg gtggtggtta
     4321 tgatttccac tacatcttgt catccatgta taatccagct gcaattaaaa acatattgat
     4381 cagaaacaac tcttttattt cttttaattt cgcacatgat ggtgtcaagt tctctgttaa
     4441 ggacagttac tcatttttgt tgtgtagttt ggccaacgct tctaaagcat ttttaaacga
     4501 agaaactttt aagaaaactg atttccctca tcacgacttg aaaactgccg atgacttata
     4561 taaggtctac aaggaatgga gttctgttaa cacagaaatt aaccatgtcg ttgaaaagga
     4621 aaagttgttg atcacctctg aacacatcgt taacttcacc aagaacgata agagtaagac
     4681 tttgatcgaa tggtctaagg attactgtag aaacgacgta ttggtcttgt ctaaggtttg
     4741 gttggagttt aaaaacgctg tagaagatat ttttaactgc gaattggtag accaaaccat
     4801 gactttggct ggtttgtcat acaagttgtt ccaagcaaac atgccattcg atgtcgaatt
     4861 gagacatcct aataaggaag actacttcaa catgagagaa gcattgattg gtggtagatg
     4921 tatctctgtt aacggtatct acaaggatgt tttgtgctta gacgtaaaat cattatatcc
     4981 agcttctatg gctttctatg atcaacctta cggttctttt aagagagttt catccagacc
     5041 aaaagatgaa ttgggtatct attacgttag agtaacccca aacagaaaca acaagtcaaa
     5101 ctttttccct atcagatccc ataataagat cacttacaac aacttcgaag aatcaactta
     5161 tattgcttgg tacacaaacg ttgatatcga cattggtttg tcagaaggtc acaacatcga
     5221 atacatccct ttcgattcct atggtaacat tggttactca tggtccaaaa agggtaaaat
     5281 ttttgaaaag tacatcaagg atgttttgta caaattgaag ataaaatatg aaaagcaaaa
     5341 taacaaggta aaaagaaatg tcatcaaaat catcatgaac tcattgtggg gtaaattcgc
     5401 ccaaaagtgg gttaacttcg aatacttcat caagtcagaa gatgacattg atttcgaatc
     5461 cgaagaagca tataaaattt gggatacaga cttcatgttg atcaaaaaga ttaaggaatc
     5521 tacctacagt tctaagccaa tacaaaacgg tgtttttact ttaagttggg ccagatatca
     5581 catgaagtct atttgggatg caggtgccaa agaaggtgct gaatgtatct attcagatac
     5641 agactccata ttcgttcata aggaacactt caaaaagaat gcaaagttta tgttgaacgg
     5701 tttgaaagta ccaatcatcg gttctgaagt cggtcaattg gaattagaat gtgaatttga
     5761 taaattgttg tgcgccggta aaaagcaata catgggtttc tacacttact tccaagatgg
     5821 taaaccttgc atcaaggaaa agaaaagatt caaaggtata ccaagtaact acatcatacc
     5881 tgaattgtat gctcacttgt tatctggtgc agataaggaa gccaaaattc aatttttaaa
     5941 attcagaaga gaatggggtt ccgtcaaggg ttacatagaa aataagactg tcaaagccac
     6001 ctaactcgag gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     6061 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
     6121 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcacta
     6181 gtccatggac gcgtggtacc ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgactgatcc ttcaactcag
     6901 caaaagttcg atttattcaa caaagccacg ttgtgtctca aaatctctga tgttacattg
     6961 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     7021 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctccaggccg cgattaaatt
     7081 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     7141 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     7201 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     7261 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     7321 tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     7381 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     7441 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     7501 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     7561 aagtctggaa agaaatgcat aagcttttgc cattctcacc gg
//
