LOCUS       sequence-376466-        5440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  untagged human PCNA with a hypomorphic disease-associated serine to 
            isoleucine mutation at residue 228.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5440)
  AUTHORS   Duffy CM, Hilbert BJ, Kelch BA
  TITLE     A Disease-Causing Variant in PCNA Disrupts a Promiscuous Protein 
            Binding Site.
  JOURNAL   J Mol Biol. 2016 Mar 27;428(6):1023-1040. doi: 
            10.1016/j.jmb.2015.11.029. Epub 2015 Dec 11.
  PUBMED    26688547
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"; doi: 
            "10.1016/j.jmb.2015.11.029"; journalName: "J Mol Biol"; date: 
            "2016-03-27- 27"; volume: "428"; issue: "6"; pages: "1023-1040"
COMMENT     Sequence Label: hPCNA-S228I
FEATURES             Location/Qualifiers
     source          1..5440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    197..336
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(262..281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     422..444
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(438..629)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2087..2106
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2190..2209
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2190..2208
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2240..2262
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3159..3177)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3173..3220
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3586..3591,3592..3608,3609..3614))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3641..3659)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3727..3831
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3832..3900,3901..4692)
                     /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(4050..4069)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4863..5440,1..11)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5352..5371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
       61 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      121 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      181 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
      241 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
      301 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
      361 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
      421 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
      481 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
      541 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
      601 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
      661 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
      721 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
      781 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
      841 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
      901 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
      961 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     1021 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     1081 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg
     1141 gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc
     1201 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
     1261 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
     1321 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
     1381 cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga
     1441 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
     1501 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
     1561 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
     1621 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     1681 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     1741 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     1801 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     1861 ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc cttatgcgac tcctgcatta
     1921 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     1981 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     2041 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     2101 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     2161 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg
     2221 gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata tgttcgaggc
     2281 gcgcctggtc cagggctcca tcctcaagaa ggtgttggag gcactcaagg acctcatcaa
     2341 cgaggcctgc tgggatatta gctccagcgg tgtaaacctg cagagcatgg actcgtccca
     2401 cgtctctttg gtgcagctca ccctgcggtc tgagggcttc gacacctacc gctgcgaccg
     2461 caacctggcc atgggcgtga acctcaccag tatgtccaaa atactaaaat gcgccggcaa
     2521 tgaagatatc attacactaa gggccgaaga taacgcggat accttggcgc tagtatttga
     2581 agcaccaaac caggagaaag tttcagacta tgaaatgaag ttgatggatt tagatgttga
     2641 acaacttgga attccagaac aggagtacag ctgtgtagta aagatgcctt ctggtgaatt
     2701 tgcacgtata tgccgagatc tcagccatat tggagatgct gttgtaattt cctgtgcaaa
     2761 agacggagtg aaattttctg caagtggaga acttggaaat ggaaacatta aattgtcaca
     2821 gacaagtaat gtcgataaag aggaggaagc tgttaccata gagatgaatg aaccagttca
     2881 actaactttt gcactgaggt acctgaactt ctttacaaaa gccactccac tctcttcaac
     2941 ggtgacactc attatgtctg cagatgtacc ccttgttgta gagtataaaa ttgcggatat
     3001 gggacactta aaatactact tggctcccaa gatcgaggat gaagaaggat cttaggcatt
     3061 cttaaaattc aagaaaataa aactaagctc tttgagaact gcttctaagg atccggctgc
     3121 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     3181 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     3241 cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta
     3301 gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg
     3361 cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc
     3421 tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
     3481 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
     3541 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga
     3601 tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt
     3661 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3721 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3781 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3841 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3901 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3961 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4021 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     4081 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4141 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4201 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4261 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4321 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     4381 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4441 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4501 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4561 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4621 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4681 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4741 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4801 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4861 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4921 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4981 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5041 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5101 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5161 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5221 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5281 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5341 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5401 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
//
