LOCUS       sequence-465754-        4581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing the Insert 2 to be assembled in POC1355.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4581)
  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 4581)
  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: POC1344
FEATURES             Location/Qualifiers
     source          1..4581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    353..377
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(533..1681)
                     /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(2017..2605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2097..2116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2776..3570)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(3571..3675)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3762..4393
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4485..4509
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cactctgtga cttggtctca gcaaggggtt ctacgtggct agttaaatag gtgcgtgatg
       61 caagatttat actagatttt tcaaatttgg tccgtatctt ggaaacatgg ggctggccgt
      121 ccactatctt ggtatgcggt cgttgtactg ggaaattaag atgccattgt tggtactcac
      181 gtgtgtatat gaggctgttt gtatccatgt agtgacggtg gagtaacaag atgttacgaa
      241 gtaggaattt cggggtaggt atcgaaggat tagatcattc aggccttctt gcatccggct
      301 cagcctgtgt actatgagac caagtcacga agtggtttaa actatcagtg tttgacagga
      361 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
      421 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
      481 cagatcaggc gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt
      541 tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact
      601 cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc
      661 acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt
      721 cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt
      781 agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg
      841 ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc
      901 caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg
      961 ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg
     1021 tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca
     1081 ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt
     1141 tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg
     1201 tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg
     1261 caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg
     1321 cggcctgctt ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct
     1381 tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct
     1441 gttcaagacg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca
     1501 gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact
     1561 ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg
     1621 cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca
     1681 ttcaccctcc ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat
     1741 ttgacccgcc tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc
     1801 cgtagaccgt ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata
     1861 ccagcgaccc cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga
     1921 tgcggaagaa gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc
     1981 tgccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2701 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2761 aacttggtct gacagctaaa acaattcatc cagtaaaata taatatttta ttttctccca
     2821 atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc
     2881 ctccctgagc gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct
     2941 cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag
     3001 gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag
     3061 ctcgcgcgga tctttaaatg gagtgtcctc ttcccagttt tcgcaatcca catcggccag
     3121 atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg
     3181 acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat
     3241 cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact
     3301 catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag
     3361 ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt
     3421 ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac
     3481 cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt
     3541 caattccggt gatattctca ttttagccat actcttcctt tttcaatatt attgaagcat
     3601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3661 aataggggtt ccgcgcacga attggccagc gctgccattt ttggggtgag gccgttcgcg
     3721 gccgaggggc gcagcccctg gggggatggg aggcccgcgt tagcgggccg ggagggttcg
     3781 agaagggggg gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt
     3841 aaaaacaagg tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc
     3901 gaaaaacggg cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa
     3961 tgtcaatagg tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc
     4021 ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag
     4081 gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga
     4141 ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc
     4201 gccgggtgag tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa
     4261 gttttccgcg aggtatccac aacgccggcg gccgcggtgt ctcgcacacg gcttcgacgg
     4321 cgtttctggc gcgtttgcag ggccatagac ggccgccagc ccagcggcga gggcaaccag
     4381 cccggtgagc gtcgcaaagg agatcctgat ctgactgatg ggctgcctgt atcgagtggt
     4441 gattttgtgc cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt
     4501 ggtgtaaaca aattgacgct tagacaactt aataacacat tgcggacgtt tttaatgtac
     4561 tggggtggat gcagtgggcc c
//
