LOCUS       sequence-202078-        3528 bp    DNA     circular SYN 15-APR-2025
DEFINITION  M. sedula PCNA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3528)
  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 3528)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-MsePCNA1
FEATURES             Location/Qualifiers
     source          1..3528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..40
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(918..935)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(918..934)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(927..949)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1142..1161)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1261..1283
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1321..1339)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1407..1511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1512..1580,1581..2372)
                     /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(1730..1749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2543..3131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3032..3051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3285..3302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3419..3440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3455..3460,3461..3478,3479..3485)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3493..3509
                     /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     3498..3520
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3517..3528,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3517..3528,1..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 acagacgcgg agcaacccag aaacgaatgc ctgcgccctt ttccagcggc gcagtcagtt
      181 tcatcggaat acccgcaccg aaaccgatct caatctcttt ggagaaggac agtgcgctgc
      241 taactttctc cagaatcgcc agggagtaac ggctggagcc ctgtttctca atttccagtt
      301 tagtcagcgg tttaccgttc ttcagcaccg cggagtattt cttgccagct tcctcaacct
      361 cgaagctcac gtggtcatcc tccgctttga tttccacttc ctcgccaacc tgcatggatt
      421 catccaccag gatgctcagg atgtcaccgc caatacccat agtaacagag tgcgcaacag
      481 acggctcttt cagaatctgc ggttcacctt tttccgcctt gatggacaga gaggagcggg
      541 tgccggtttt ctcgtccttc atgctgatgc ggatgccttc gttggtttca gtaatctcca
      601 cggagctgcg gcccttcatc ttggacagga tctttttcag ttcgcccagg ttcagcttca
      661 cagaaatcgg cttctcgata gagtattctt ccagagcgtc tttaccaatt tcaaccacac
      721 ccatcataac cttgtcatcg gtcagatact tggagccgat gccctcttca gtgaagttca
      781 gggtcagctc gtcgctgatt ttgctaatgc tagacacgat gtaatagaag tcacgagagc
      841 taccatagat tgcacggaac atcatatggg attcccatgg catgccaatc ggatccccgg
      901 gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
      961 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     1021 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggccctga
     1081 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatag tgcactctca
     1141 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     1201 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     1261 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     1321 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     1381 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1441 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1501 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1561 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1621 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1681 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1741 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1801 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1861 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1921 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1981 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2041 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2101 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2161 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2221 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2281 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2341 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2401 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2461 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2521 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2581 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2641 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2701 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2761 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2821 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2881 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2941 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3001 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3061 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3241 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3301 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3361 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3421 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     3481 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagct
//
