LOCUS       sequence-97872-e        8763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to express human Pol iota recoded to yeast codons in yeast .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8763)
  AUTHORS   Makarova AV, Grabow C, Gening LV, Tarantul VZ, Tahirov TH, Bessho T,
            Pavlov YI
  TITLE     Inaccurate DNA synthesis in cell extracts of yeast producing active 
            human DNA polymerase iota.
  JOURNAL   PLoS One. 2011 Jan 31;6(1):e16612. doi: 
            10.1371/journal.pone.0016612.
  PUBMED    21304950
REFERENCE   2  (bases 1 to 8763)
  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: "PLoS One.";
            date: "2011-01-31"; volume: "6(1):e16612. doi"; pages: " 
            10.1371/journal.pone.0016612"
COMMENT     Sequence Label: pESC-URA-hPOLISc
FEATURES             Location/Qualifiers
     source          1..8763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        196..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /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(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(1885..2050)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     1971..1995
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2198..2221)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        2268..2932
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    2484..2601
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2896..2919
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     2943..2962
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2943..2961
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2979..3008
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4820..4831
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      5170..5359
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5431..5448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5602..6190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5682..5701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6361..7152,7153..7221))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6984..7003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7222..7326)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7353..8695
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(8316..8363)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     8736..8754
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1861 ttcgctatta cgccagctga attggagcga cctcatgcta tacctgagaa agcaacctga
     1921 cctacaggaa agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt
     1981 aaaatttgta tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata
     2041 agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc aacgatttga cccttttcca
     2101 tcttttcgta aatttctggc aaggtagaca agccgacaac cttgattgga gacttgacca
     2161 aacctctggc gaagaattgt taattaagag ctcagatctt atcgtcgtca tccttgtaat
     2221 ccatcgatac tagtgcggcc gccctttagt gagggttgaa ttcgaatttt caaaaattct
     2281 tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc
     2341 atatacatat ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa
     2401 gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa
     2461 ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggtgac agccctccga
     2521 aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt
     2581 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg
     2641 aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa
     2701 caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc
     2761 gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt
     2821 aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat
     2881 caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa accccggatc
     2941 cgtaatacga ctcactatag ggcccgggcg tcgacatgga acagaagttg atttccgaag
     3001 aagacctcga gatggaattg gctgacgttg gtgctgctgc ttcttctcaa ggtgttcacg
     3061 accaagtttt gccaacccca aacgcttctt ctagagttat cgttcacgtt gacttggact
     3121 gtttctacgc tcaagttgaa atgatctcta acccagaatt gaaggacaag ccattgggtg
     3181 ttcaacaaaa gtacttggtt gttacctgta actacgaagc tagaaagttg ggtgttaaga
     3241 agttgatgaa cgttagagac gctaaggaaa agtgtccaca attggttttg gttaacggtg
     3301 aagacttgac cagatacaga gaaatgtctt acaaggttac cgaattgttg gaagaattct
     3361 ctccagttgt tgaaagattg ggtttcgacg aaaacttcgt tgacttgacc gaaatggttg
     3421 aaaagagatt gcaacaattg caatctgacg aattgtctgc tgttaccgtt tctggtcacg
     3481 tttacaacaa ccaatctatc aacttgttgg acgttttgca catcagattg ttggttggtt
     3541 ctcaaatcgc tgctgaaatg agagaagcta tgtacaacca attgggtttg accggttgtg
     3601 ctggtgttgc ttctaacaag ttgttggcta agttggtttc tggtgttttc aagccaaacc
     3661 aacaaaccgt tttgttgcca gaatcttgtc aacacttgat ccactctttg aaccacatca
     3721 aggaaatccc aggtatcggt tacaagaccg ctaagtgttt ggaagctttg ggtatcaact
     3781 ctgttagaga cttgcaaacc ttctctccaa agatcttgga aaaggaattg ggtatctctg
     3841 ttgctcaaag aatccaaaag ttgtctttcg gtgaagacaa ctctccagtt atcttgtctg
     3901 gtccaccaca atctttctct gaagaagact ctttcaagaa gtgttcttct gaagttgaag
     3961 ctaagaacaa gatcgaagaa ttgttggctt ctttgttgaa cagagtttgt caagacggta
     4021 gaaagccaca caccgttaga ttgatcatca gaagatactc ttctgaaaag cactacggta
     4081 gagaatctag acaatgtcca atcccatctc acgttatcca aaagttgggt accggtaact
     4141 acgacgttat gaccccaatg gttgacatct tgatgaagtt gttcagaaac atggttaacg
     4201 ttaagatgcc attccacttg accttgttgt ctgtttgttt ctgtaacttg aaggctttga
     4261 acaccgctaa gaagggtttg atcgactact acttgatgcc atctttgtct accacctcta
     4321 gatctggtaa gcactctttc aagatgaagg acacccacat ggaagacttc ccaaaggaca
     4381 aggaaaccaa cagagacttc ttgccatctg gtagaatcga atctaccaga accagagaat
     4441 ctccattgga caccaccaac ttctctaagg aaaaggacat caacgaattc ccattgtgtt
     4501 ctttgccaga aggtgttgac caagaagttt tcaagcaatt gccagttgac atccaagaag
     4561 aaatcttgtc tggtaagtct agagaaaagt tccaaggtaa gggttctgtt tcttgtccat
     4621 tgcacgcttc tagaggtgtt ttgtctttct tctctaagaa gcaaatgcaa gacatcccaa
     4681 tcaacccaag agaccacttg tcttcttcta agcaagtttc ttctgtttct ccatgtgaac
     4741 caggtacctc tggtttcaac tcttcttctt cttcttacat gtcttctcaa aaggactact
     4801 cttactactt ggacaacaga ttgaaggacg aaagaatctc tcaaggtcca aaggaaccac
     4861 aaggtttcca cttcaccaac tctaacccag ctgtttctgc tttccactct ttcccaaact
     4921 tgcaatctga acaattgttc tctagaaacc acaccaccga ctctcacaag caaaccgttg
     4981 ctaccgactc tcacgaaggt ttgaccgaaa acagagaacc agactctgtt gacgaaaaga
     5041 tcaccttccc atctgacatc gacccacaag ttttctacga attgccagaa gctgttcaaa
     5101 aggaattgtt ggctgaatgg aagagaaccg gttctgactt ccacatcggt cacaagtaag
     5161 ctagctaaga tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     5221 tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     5281 tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     5341 ggttttggga cgctcgaaga tccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5761 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6001 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6121 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6241 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6301 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6361 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6421 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6481 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6541 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6601 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6661 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6721 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6781 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6841 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6901 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6961 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7021 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7081 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7141 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7201 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7261 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7321 tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt cattttgtag
     7381 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     7441 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     7501 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     7561 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     7621 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     7681 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     7741 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     7801 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     7861 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     7921 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     7981 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     8041 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     8101 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     8161 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     8221 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     8281 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat
     8341 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     8401 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     8461 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     8521 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     8581 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     8641 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatact
     8701 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8761 gtc
//
