LOCUS       sequence-191858-        5816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative vector for expression of His-tagged POL30 (PCNA) 
            mutant K164R; Leu2 marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5816)
  AUTHORS   Davies AA, Ulrich HD
  TITLE     Detection of PCNA modifications in Saccharomyces cerevisiae.
  JOURNAL   Methods Mol Biol. 2012;920:543-67. doi: 
            10.1007/978-1-61779-998-3_36.
  PUBMED    22941627
REFERENCE   2  (bases 1 to 5816)
  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: "Methods Mol
            Biol."; date: "2012"; volume: "920:543-67. doi"; pages: " 
            10.1007/978-1-61779-998-3_36"
COMMENT     Sequence Label: YIp128-P30-His-pol30(K164R)
FEATURES             Location/Qualifiers
     source          1..5816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             576..593
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1401..1588
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1456..1480)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1612..1628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1612..1628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1625..1647)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1636..1652
                     /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(1660..1666,1667..1684,1685..1690))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1705..1726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1843..1860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2014..2602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2094..2113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2773..3564,3565..3633))
                     /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     3396..3415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3634..3738)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3806..3824
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3826..4231
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             4232..5326
                     /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"
     primer_bind     complement(5467..5489)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5589..5608
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(5802..5816,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5816,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttcgccctta ttccttaact gctgaagaaa agcaacccta
       61 tgaatctaag gctcaagcag acaagaagag atacgaatct gaaaaggaat tgtacaatgc
      121 tacacgtgct tgatctaaca actcaatgtt ctctacctcc catacaacgt acttttgaaa
      181 cttcagaaag aagttgagta ggttgcatta tgtgttatta ttattactat tatcattatt
      241 ttacttatat atccttcttt tcttttatgt attctcttcc tttttcgcac aacttatgct
      301 gattcttcaa tatctaatta tttagcattt ttcttctcca tccgacgcac caaaacgcga
      361 aacgcgtaac tttttttttg gatttcaact gatagttttc gtactttgct tcctctggta
      421 cataaaatta tatataagaa acacttttgc tttagccttc ctttctttcc acttgcacct
      481 ttcactttcg ccgtcctttt tcactcacag caacaagcag caagcactaa gtacgcagtc
      541 aaaagagaga aaaaggatcc aatatgagag gatctcacca tcaccatcac catggcccgg
      601 gaatgttaga agcaaaattt gaagaagcat cccttttcaa gagaataatt gatggtttca
      661 aagattgtgt ccagttggtc aatttccaat gtaaagaaga tggtatcatt gcacaagctg
      721 tcgatgactc aagagttcta ttggtctcct tggaaatagg tgtcgaagcc ttccaagaat
      781 atagatgtga ccatcctgtt acgttaggta tggatctaac ctcactaagt aaaatcctac
      841 gttgtggtaa caacaccgat acattaacac taattgctga caacacaccg gattccatca
      901 tcttattatt tgaggatacc aagaaagacc gtatagccga atactctctg aaattgatgg
      961 atatcgatgc tgatttctta aagattgaag aattacagta cgactccacc ctgtcattgc
     1021 catcttccga attctctaaa attgttcgtg acttgtccca attgagtgat tctattaata
     1081 tcatgataac cagagaaaca ataaagtttg tagctgacgg tgatatcgga tcaggttcag
     1141 tcataataaa accattcgtg gatatggaac atcctgaaac aagcatcaaa cttgaaatgg
     1201 atcaacctgt cgacttgacg ttcggagcta aatatttatt ggacatcatt aagggctcct
     1261 ccctttctga tagagttggt atcaggctct ccagcgaagc tcctgcttta ttccaatttg
     1321 atttgaagag tgggttccta cagtttttct tggctcctaa atttaatgac gaagaataac
     1381 tgcagccaag ctaattccgg gcgaatttct tatgatttat gatttttatt attaaataag
     1441 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     1501 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
     1561 tcgctcttat tgaccacacc tctaccggca tgcaagcttg gcgtaatcat ggtcatagct
     1621 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1681 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1741 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1801 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1861 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1921 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1981 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2641 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2701 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2761 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2821 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2881 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2941 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3001 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3061 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3121 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3181 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3241 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3301 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3361 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3421 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3481 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3541 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3601 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3661 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3721 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     3781 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccaactg tgggaatact
     3841 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
     3901 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
     3961 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
     4021 ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa
     4081 atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt gttttttcca
     4141 ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata
     4201 tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga tcgtcgtttt
     4261 gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc ttaaagctat
     4321 ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg gtggtgctgc
     4381 tatcgatgct acaggtgttc cacttccaga tgaggcgctg gaagcctcca agaaggctga
     4441 tgccgttttg ttaggtgctg tgggtggtcc taaatggggt acaggtagtg ttagacctga
     4501 acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa gaccatgtaa
     4561 ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg ctaaaggtac
     4621 tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga gaaaggaaga
     4681 cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag tgcaaagaat
     4741 cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta tttggtcctt
     4801 ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg aggaaaccat
     4861 caagaacgaa tttcctacat tgaaggttca acatcaattg attgattctg ccgccatgat
     4921 cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca tgtttggtga
     4981 tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc catctgcgtc
     5041 cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat gccacggttc
     5101 tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat
     5161 gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag atgcagttaa
     5221 aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca gtaccaccga
     5281 agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag attctctttt
     5341 tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg acacgaaatt
     5401 acaaaatgga atatgttcat agggtaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5461 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5521 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5581 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5641 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5701 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5761 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgtt
//
