LOCUS       sequence-204679-        3519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M. sedula PCNA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3519)
  AUTHORS   Iwata F, Hirakawa H, Nagamune T
  TITLE     Three proliferating cell nuclear antigen homologues from 
            Metallosphaera sedula form a head-to-tail heterotrimer.
  JOURNAL   Sci Rep. 2016 May 27;6:26588. doi: 10.1038/srep26588.
  PUBMED    27228945
REFERENCE   2  (bases 1 to 3519)
  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: "Sci Rep."; 
            date: "2016-05-27"; volume: "6:26588. doi"; pages: " 
            10.1038/srep26588"
COMMENT     Sequence Label: pMD-MsePCNA2
FEATURES             Location/Qualifiers
     source          1..3519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..40
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(909..926)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(909..925)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(918..940)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1133..1152)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1252..1274
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1312..1330)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1398..1502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1503..1571,1572..2363)
                     /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     complement(1721..1740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2534..3122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3023..3042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3276..3293
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3410..3431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3446..3451,3452..3469,3470..3476)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3484..3500
                     /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)."
     primer_bind     3489..3511
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3508..3519,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3508..3519,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag ctatttaggt gacactatag gggaaagctt gcatgcctgc
       61 aggtcgactc tagaggatct actagtcata tggattcatg ccatggcatg ccggatcccg
      121 attccgcacg cggtgcaata tagaagtcgc cgtagccttc ctgcggcagt tcgaaacgca
      181 gcttaatcgg cagctgggaa ccaaacatca gttcaacgat gtcggaagag ttgcgcatct
      241 tcgcggtctt aaccaggtag tccatgccgt aggtgccctt agcgtccgca ccggtcgctt
      301 ccagcagagt accggattct tcggacagct ccaccttgct agaaccaacg tcgccctcaa
      361 ccatgaaagt cagtttacca ccctccgcag acagggtcag cgcgtcaccc aggtcgccca
      421 ggatctgcat aacgttggaa aaggtagagg tcagcatttt cgccttgaac gcgaactcca
      481 gtttgatgga cggcagagac ggttcctcgc ccatcaggat cggcagataa aaggtgcgct
      541 caaagtcacc gtccagagtg atcatcagac ggtcctcacc agactggaag gtcagagagt
      601 cgttcttctt cacgtttttc aggctcgcga tgatctcctc cagcttcacg gtaacggttt
      661 ctttctctgc gctttcgtac tcgtggaagt acgcctgcgg gatgaagatg tcgatcagcg
      721 caacacgtgc cgggtcaaca ccgctcagac gaacaccttc tttggtaccg atcagggtaa
      781 tctccggcat aaactcgccg atggtacgga agatggtcgc catagagttc gcgtcgataa
      841 ctttgaattt catcatatgg gattcccatg gcatgccaat cggatccccg ggtaccgagc
      901 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      961 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     1021 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggccctg atgcggtatt
     1081 ttctccttac gcatctgtgc ggtatttcac accgcatata gtgcactctc agtacaatct
     1141 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     1201 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1261 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     1321 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1381 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1441 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1501 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1561 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1621 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1681 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1741 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1801 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     1861 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     1921 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     1981 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2041 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2101 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2161 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2221 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2281 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2341 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2401 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2461 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2521 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2581 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2641 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2701 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2761 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2821 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2881 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2941 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3001 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3061 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3121 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3181 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3241 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3301 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3361 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     3421 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     3481 gaattgtgag cggataacaa tttcacacag gaaacagct
//
