LOCUS       sequence-374280-        5440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  untagged human PCNA with a cysteine to serine mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5440)
  AUTHORS   Magrino J, Munford V, Martins DJ, Homma TK, Page B, Gaubitz C, 
            Freire BL, Lerario AM, Vilar JB, Amorin A, Leao EKE, Kok F, Menck 
            CF, Jorge AA, Kelch BA
  TITLE     A thermosensitive PCNA allele underlies an 
            ataxia-telangiectasia-like disorder.
  JOURNAL   J Biol Chem. 2023 May;299(5):104656. doi: 10.1016/j.jbc.2023.104656.
            Epub 2023 Mar 27.
  PUBMED    36990216
REFERENCE   2  (bases 1 to 5440)
  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: "J Biol 
            Chem."; date: "2023-05"; volume: "299(5):104656. doi"; pages: " 
            10.1016/j.jbc.2023.104656. Epub 2023 Mar 27"
COMMENT     Sequence Label: hPCNA-C148S
FEATURES             Location/Qualifiers
     source          1..5440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(36..55)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     196..218
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(212..403)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1861..1880
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1964..1983
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1964..1982
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2014..2036
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2933..2951)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2947..2994
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3360..3365,3366..3382,3383..3388))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3415..3433)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3501..3605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3606..3674,3675..4466)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3824..3843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4637..5225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5126..5145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5379..5396
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5411..5440,1..110)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
       61 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
      121 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
      181 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
      241 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
      301 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
      361 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
      421 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
      481 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
      541 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
      601 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
      661 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
      721 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
      781 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
      841 acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc
      901 gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg
      961 cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag
     1021 cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa
     1081 cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt
     1141 gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct
     1201 ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga
     1261 cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg
     1321 gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc
     1381 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     1441 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     1501 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     1561 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     1621 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgacgct ctcccttatg
     1681 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     1741 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     1801 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     1861 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     1921 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
     1981 gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata
     2041 catatgttcg aggcgcgcct ggtccagggc tccatcctca agaaggtgtt ggaggcactc
     2101 aaggacctca tcaacgaggc ctgctgggat attagctcca gcggtgtaaa cctgcagagc
     2161 atggactcgt cccacgtctc tttggtgcag ctcaccctgc ggtctgaggg cttcgacacc
     2221 taccgctgcg accgcaacct ggccatgggc gtgaacctca ccagtatgtc caaaatacta
     2281 aaatgcgccg gcaatgaaga tatcattaca ctaagggccg aagataacgc ggataccttg
     2341 gcgctagtat ttgaagcacc aaaccaggag aaagtttcag actatgaaat gaagttgatg
     2401 gatttagatg ttgaacaact tggaattcca gaacaggagt acagctgtgt agtaaagatg
     2461 ccttctggtg aatttgcacg tatatcccga gatctcagcc atattggaga tgctgttgta
     2521 atttcctgtg caaaagacgg agtgaaattt tctgcaagtg gagaacttgg aaatggaaac
     2581 attaaattgt cacagacaag taatgtcgat aaagaggagg aagctgttac catagagatg
     2641 aatgaaccag ttcaactaac ttttgcactg aggtacctga acttctttac aaaagccact
     2701 ccactctctt caacggtgac actcagtatg tctgcagatg taccccttgt tgtagagtat
     2761 aaaattgcgg atatgggaca cttaaaatac tacttggctc ccaagatcga ggatgaagaa
     2821 ggatcttagg cattcttaaa attcaagaaa ataaaactaa gctctttgag aactgcttct
     2881 aaggatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     2941 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     3001 ggaggaacta tatccggata tccacaggac gggtgtggtc gccatgatcg cgtagtcgat
     3061 agtggctcca agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc
     3121 tccgagaacg ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata
     3181 gtgactggcg atgctgtcgg aatggacgat atcccgcaag aggcccggca gtaccggcat
     3241 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
     3301 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
     3361 ttaaagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
     3421 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3601 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3661 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3721 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3781 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3841 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3901 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3961 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4021 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4081 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4141 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4201 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4261 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4321 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4381 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4441 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5341 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5401 cggaagagcg cctgatgcgg tattttctcc ttacgcatct
//
