LOCUS       sequence-182200-        5822 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 5822)
  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 5822)
  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..5822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(942..1733,1734..1802))
                     /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     1565..1584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1975..1993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1995..2399
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2412..3506
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(3647..3669)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3769..3788
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3982..4004
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3996..4013
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3997..4013
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4564..4581
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5392..5579
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5447..5471)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5603..5619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5603..5619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5616..5638)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5627..5643
                     /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(5651..5657,5658..5675,5676..5681))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5696..5717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccaactgt gggaatactc aggtatcgta agatgcaaga gttcgaatct
     2041 cttagcaacc attatttttt tcctcaacat aacgagaaca cacaggggcg ctatcgcaca
     2101 gaatcaaatt cgatgactgg aaattttttg ttaatttcag aggtcgcctg acgcatatac
     2161 ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag aggccggaac cggcttttca
     2221 tatagaatag agaagcgttc atgactaaat gcttgcatca caatacttga agttgacaat
     2281 attatttaag gacctattgt tttttccaat aggtggttag caatcgtctt actttctaac
     2341 ttttcttacc ttttacattt cagcaatata tatatatatt tcaaggatat accattctaa
     2401 tgtctgcccc tatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac cacgttggtc
     2461 aagaaatcac agccgaagcc attaaggttc ttaaagctat ttctgatgtt cgttccaatg
     2521 tcaagttcga tttcgaaaat catttaattg gtggtgctgc tatcgatgct acaggtgtcc
     2581 cacttccaga tgaggcgctg gaagcctcca agaaggttga tgccgttttg ttaggtgctg
     2641 tgggtggtcc taaatggggt acaggtagtg ttagacctga acaaggttta ctaaaaatcc
     2701 gtaaagaact tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc gactctcttt
     2761 tagacttatc tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt gttgtcagag
     2821 aattagtggg aggtatttac tttggtaaga gaaaggaaga cgatggtgat ggtgtcgctt
     2881 gggatagtga acaatacacc gttccagaag tgcaaagaat cacaagaatg gccgctttca
     2941 tggccctaca acatgagcca ccattgccta tttggtcctt ggataaagct aatgttttgg
     3001 cctcttcaag attatggaga aaaactgtgg aggaaaccat caagaacgaa tttcctacat
     3061 tgaaggttca acatcaattg attgattctg ccgccatgat cctagttaag aacccaaccc
     3121 acctaaatgg tattataatc accagcaaca tgtttggtga tatcatctcc gatgaagcct
     3181 ccgttatccc aggttccttg ggtttgttgc catctgcgtc cttggcctct ttgccagaca
     3241 agaacaccgc atttggtttg tacgaaccat gccacggttc tgctccagat ttgccaaaga
     3301 ataaggttga ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa ttgtcattga
     3361 acttgcctga agaaggtaag gccattgaag atgcagttaa aaaggttttg gatgcaggta
     3421 tcagaactgg tgatttaggt ggttccaaca gtaccaccga agtcggtgat gctgtcgccg
     3481 aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttatgatat ttgtacataa
     3541 actttataaa tgaaattcat aatagaaacg acacgaaatt acaaaatgga atatgttcat
     3601 agggtaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3661 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3721 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     3781 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3841 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     3901 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     3961 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcc
     4021 ttaactgctg aagaaaagca accctatgaa tctaaggctc aagcagacaa gaagagatac
     4081 gaatctgaaa aggaattgta caatgctaca cgtgcttgat ctaacaactc aatgttctct
     4141 acctcccata caacgtactt ttgaaacttc agaaagaagt tgagtaggtt gcattatgtg
     4201 ttattattat tactattatc attattttac ttatatatcc ttcttttctt ttatgtattc
     4261 tcttcctttt tcgcacaact tatgctgatt cttcaatatc taattattta gcatttttct
     4321 tctccatccg acgcaccaaa acgcgaaacg cgtaactttt tttttttgga tttcaactga
     4381 tagttttcgt actttgcttc ctctggtaca taaaattata tataagaaac acttttgctt
     4441 tagccttcct ttctttccac ttgcaccttt cactttcgcc gtcctttttc actcacagca
     4501 acaagcagca agcactaagt acgcagtcaa aagagagaaa aaggatccaa tatgagagga
     4561 tctcaccatc accatcacca tggcccggga aaaatgttag aagcaaaatt tgaagaagca
     4621 tcccttttca agagaataat tgatggtttc aaagattgtg tccagttggt caatttccaa
     4681 tgtaaagaag atggtatcat tgcacaagct gtcgatgact caagagttct attggtctcc
     4741 ttggaaatag gtgtcgaagc cttccaagaa tatagatgtg accatcctgt tacgttaggt
     4801 atggatctaa cctcactaag taaaatccta cgttgtggta acaacaccga tacattaaca
     4861 ctaattgctg acaacacacc ggattccatc atcttattat ttgaggatac caagaaagac
     4921 cgtatagccg aatactctct gaaattgatg gatatcgatg ctgatttctt aaagattgaa
     4981 gaattacagt acgactccac cctgtcattg ccatcttccg aattctctaa aattgttcgt
     5041 gacttgtccc aattgagtga ttctattaat atcatgataa ccagagaaac aataaagttt
     5101 gtagctgacg gtgatatcgg atcaggttca gtcataataa aaccattcgt ggatatggaa
     5161 catcctgaaa caagcatcaa acttgaaatg gatcaacctg tcgacttgac gttcggagct
     5221 aaatatttat tggacatcat taagggctcc tccctttctg atagagttgg tatcaggctc
     5281 tccagcgaag ctcctgcttt attccaattt gatttgaaga gtgggttcct acagtttttc
     5341 ttggctccta aatttaatga cgaagaataa ctgcagccaa gctaattccg ggcgaatttc
     5401 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     5461 taaagtgact cttaggtttt aaaacgaaaa ttcttgttct tgagtaactc tttcctgtag
     5521 gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc
     5581 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5641 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5701 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5761 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5821 gc
//
