LOCUS       sequence-465762-        4584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing the Insert 3 to be assembled in POC1355.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4584)
  AUTHORS   Flores-Fernandez CN, Lin D, Robins K, O'Callaghan CA
  TITLE     UniClo: scarless hierarchical DNA assembly without sequence 
            constraint.
  JOURNAL   Nucleic Acids Res. 2025 Jun 20;53(12):gkaf548. doi: 
            10.1093/nar/gkaf548.
  PUBMED    40548934
REFERENCE   2  (bases 1 to 4584)
  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: "Nucleic 
            Acids Res."; date: "2025-06-20"; volume: "53(12):gkaf548. doi"; 
            pages: " 10.1093/nar/gkaf548"
COMMENT     Sequence Label: POC1345
FEATURES             Location/Qualifiers
     source          1..4584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    356..380
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(536..1684)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(2020..2608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2100..2119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2779..3573)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(3574..3678)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3765..4396
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4488..4512
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cactctgtga cttggtctca actagagcgt gggaatgcaa atacgtccgt ggttgcggat
       61 ctgggccgcg gaccggtggg tgcgccttct ggtgttgggc ggtctttgag gcagacagtg
      121 aggtctagag ttggttgatc tgtagctaag acgtcacctc agagtttatc agtactgtag
      181 acttttcatt caaggtaccc ctggtcgaaa atacgcgtgg tagaatcgtt gttgcgcgta
      241 tcctttttaa gacggaagcg caccgggggg aatggtgtgg ttaagagggt tggcataacg
      301 ctaggcatgg tggttactga gaccaagtca cgaagtggtt taaactatca gtgtttgaca
      361 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
      421 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
      481 ccccagatca ggcgctggct gctgaacccc cagccggaac tgaccccaca aggccctagc
      541 gtttgcaatg caccaggtca tcattgaccc aggcgtgttc caccaggccg ctgcctcgca
      601 actcttcgca ggcttcgccg acctgctcgc gccacttctt cacgcgggtg gaatccgatc
      661 cgcacatgag gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc gagctgaaat
      721 agtcgaacat ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat atgaatttcg
      781 tgtagtggtc gccagcaaac agcacgacga tttcctcgtc gatcaggacc tggcaacggg
      841 acgttttctt gccacggtcc aggacgcgga agcggtgcag cagcgacacc gattccaggt
      901 gcccaacgcg gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac aggcattcct
      961 cggccttcgt gtaataccgg ccattgatcg accagcccag gtcctggcaa agctcgtaga
     1021 acgtgaaggt gatcggctcg ccgatagggg tgcgcttcgc gtactccaac acctgctgcc
     1081 acaccagttc gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct
     1141 tgttgacgtg gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg
     1201 tggtgaacag ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg
     1261 gcgcaatatc gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg tgtttcagca
     1321 acgcggcctg cttggcctcg ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct
     1381 tcttggtcgt catagttcct cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct
     1441 cctgttcaag acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg gcagggggag
     1501 ccagttgcac gctgtcgcgc tcgatcttgg ccgtagcttg ctggaccatc gagccgacgg
     1561 actggaaggt ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt tcggcatcct
     1621 cggcggaaaa ccccgcgtcg atcagttctt gcctgtatgc cttccggtca aacgtccgat
     1681 tcattcaccc tccttgcggg attgccccga ctcacgccgg ggcaatgtgc ccttattcct
     1741 gatttgaccc gcctggtgcc ttggtgtcca gataatccac cttatcggca atgaagtcgg
     1801 tcccgtagac cgtctggccg tccttctcgt acttggtatt ccgaatcttg ccctgcacga
     1861 ataccagcga ccccttgccc aaatacttgc cgtgggcctc ggcctgagag ccaaaacact
     1921 tgatgcggaa gaagtcggtg cgctcctgct tgtcgccggc atcgttgcgc cacatctagg
     1981 atctgccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2461 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2701 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2761 gtaaacttgg tctgacagct aaaacaattc atccagtaaa atataatatt ttattttctc
     2821 ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg acatactgtt cttccccgat
     2881 atcctccctg agcgaccgga cgcagaaggc aatgtcatac cacttgtccg ccctgccgct
     2941 tctcccaaga tcaataaagc cacttacttt gccatctttc acaaagatgt tgctgtctcc
     3001 caggtcgccg tgggaaaaga caagttcctc ttcgggcttt tccgtcttta aaaaatcata
     3061 cagctcgcgc ggatctttaa atggagtgtc ctcttcccag ttttcgcaat ccacatcggc
     3121 cagatcgtta ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc tattcgtata
     3181 gggacaatcc gatatgtcga tggagtgaaa gagcctgatg cactccgcat acagctcgat
     3241 aatcttttca gggctttgtt catcttcata ctcttccgag caaaggacgc catcggcctc
     3301 actcatgagc agattgctcc agccatcatg ccgttcaaag tgcaggacct ttggaacagg
     3361 cagctttcct tccagccata gcatcatgtc cttttcccgt tccacatcat aggtggtccc
     3421 tttataccgg ctgtccgtca tttttaaata taggttttca ttttctccca ccagcttata
     3481 taccttagca ggagacattc cttccgtatc ttttacgcag cggtattttt cgatcagttt
     3541 tttcaattcc ggtgatattc tcattttagc catactcttc ctttttcaat attattgaag
     3601 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3661 acaaataggg gttccgcgca cgaattggcc agcgctgcca tttttggggt gaggccgttc
     3721 gcggccgagg ggcgcagccc ctggggggat gggaggcccg cgttagcggg ccgggagggt
     3781 tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg cgcagccctg
     3841 gttaaaaaca aggtttataa atattggttt aaaagcaggt taaaagacag gttagcggtg
     3901 gccgaaaaac gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg acagcccctc
     3961 aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg tcaaggatcg
     4021 cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg cacttatccc
     4081 caggcttgtc cacatcatct gtgggaaact cgcgtaaaat caggcgtttt cgccgatttg
     4141 cgaggctggc cagctccacg tcgccggccg aaatcgagcc tgcccctcat ctgtcaacgc
     4201 cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt cagtgagggc
     4261 caagttttcc gcgaggtatc cacaacgccg gcggccgcgg tgtctcgcac acggcttcga
     4321 cggcgtttct ggcgcgtttg cagggccata gacggccgcc agcccagcgg cgagggcaac
     4381 cagcccggtg agcgtcgcaa aggagatcct gatctgactg atgggctgcc tgtatcgagt
     4441 ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat
     4501 tgtggtgtaa acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg
     4561 tactggggtg gatgcagtgg gccc
//
