LOCUS       sequence-204540-        3514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M. sedula PCNA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3514)
  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 3514)
  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-MsePCNA3
FEATURES             Location/Qualifiers
     source          1..3514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..930
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIGESLHACRSTLEDLLVIWIGMPWESHMMRIAY
                     ANAMDFKTVIEALSKLIDEVTFTFTSSGLDVVAVDRAHISLIKLHFPKEAFEEFDVEDQ
                     FRFGFNTQYMLKVMASAKRKEKIEMESREESEIVIRMLGEPPREFTIRNIEVPIQELPE
                     LKLDFDVKAKITSGGFKKAVSEIATVSDSVEIDATEMEIKLRSKESTEIEVEFSKEMGG
                     LQEIEVKKPSVSSYPSDYLEDVLVLTRLSGFLNLLFSEQKPLQLEFNMDNGGSVVYLLA
                     PNVGSGSGMPIGSPGTELEFTGRRFTTS"
     primer_bind     23..40
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(904..921)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(904..920)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(913..935)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1128..1147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1247..1269
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1307..1325)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1393..1497
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1498..1566,1567..2358)
                     /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(1716..1735)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2529..3117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3018..3037
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3271..3288
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3405..3426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3441..3446,3447..3464,3465..3471)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3479..3495
                     /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     3484..3506
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3503..3514,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3503..3514,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag ctatttaggt gacactatag gggaaagctt gcatgcctgc
       61 aggtcgactc tagaggatct actagtcata tggattggca tgccatggga atcccatatg
      121 atgcgtatcg catacgcgaa cgcgatggac ttcaaaaccg tgatcgaagc gctgtctaaa
      181 ctgatcgacg aagtgacctt cactttcacc agctctggtc tggacgttgt tgcggttgac
      241 cgtgcgcata tcagcctgat caagctgcac ttcccgaaag aagcgttcga agaatttgac
      301 gtggaagacc agttccgttt cggctttaac acccagtaca tgctgaaggt tatggcgtct
      361 gcgaaacgca aggagaagat tgagatggag tcccgtgaag agtccgaaat cgttatccgt
      421 atgctgggcg aaccgccgcg tgaatttacc atccgtaaca ttgaggttcc gatccaggaa
      481 ctgccggagc tgaaactgga tttcgacgtg aaggcgaaaa tcacttctgg cggcttcaag
      541 aaggctgtta gcgagattgc gaccgtttct gactctgtgg agattgacgc gaccgaaatg
      601 gagattaagc tgcgttccaa ggagtccacc gagatcgagg tggagttctc caaagaaatg
      661 ggtggtctgc aggaaattga ggtgaagaag ccgtctgttt cttcctaccc gtctgactac
      721 ctggaggacg ttctggttct gacccgcctg tctggcttcc tgaacctgct gttctctgag
      781 cagaaaccgc tgcagctgga gttcaacatg gacaacggtg gctccgtggt ttatctgctg
      841 gcgccgaacg ttggttcggg atccggcatg ccaatcggat ccccgggtac cgagctcgaa
      901 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
      961 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     1021 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg ccctgatgcg gtattttctc
     1081 cttacgcatc tgtgcggtat ttcacaccgc atatagtgca ctctcagtac aatctgctct
     1141 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     1201 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     1261 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     1321 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     1381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1441 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1501 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1561 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1621 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1681 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1741 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1801 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1861 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1921 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1981 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2041 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2101 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2161 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2221 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2281 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2341 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2401 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2461 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2521 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2581 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2641 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2701 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2761 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2821 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2881 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2941 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3001 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3061 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3121 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3181 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3241 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3301 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3361 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     3421 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     3481 gtgagcggat aacaatttca cacaggaaac agct
//
