LOCUS       sequence-465756-        4326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing the Insert 1 to be assembled in POC1355.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4326)
  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 4326)
  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: POC1343
FEATURES             Location/Qualifiers
     source          1..4326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    98..122
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(278..1426)
                     /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(1762..2350)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1842..1861)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2521..3315)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(3316..3420)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3507..4138
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4230..4254
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cactctgtga cttggtctca tgccttgtgt tgatactaat gcgttaatgt tcggggcaat
       61 gagaccaagt cacgaagtgg tttaaactat cagtgtttga caggatatat tggcgggtaa
      121 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
      181 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttccccagat caggcgctgg
      241 ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt
      301 catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc
      361 cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg
      421 tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg
      481 ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa
      541 acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt
      601 ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg
      661 tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc
      721 ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct
      781 cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt
      841 cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga
      901 ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc
      961 gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg
     1021 aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct
     1081 cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc
     1141 ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttca agacgacgcg
     1201 aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc
     1261 gctcgatctt ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg
     1321 gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt
     1381 cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg
     1441 ggattgcccc gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg
     1501 ccttggtgtc cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc
     1561 cgtccttctc gtacttggta ttccgaatct tgccctgcac gaataccagc gaccccttgc
     1621 ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg
     1681 tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa
     1741 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1801 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1861 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1921 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1981 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2041 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2101 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2161 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2221 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2281 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2341 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2401 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2461 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2521 ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc
     2581 cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc tgagcgaccg
     2641 gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa
     2701 gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa
     2761 gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt
     2821 aaatggagtg tcctcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa
     2881 gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc
     2941 gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg
     3001 ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct
     3061 ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca
     3121 tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt
     3181 catttttaaa tataggtttt cattttctcc caccagctta tataccttag caggagacat
     3241 tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat
     3301 tctcatttta gccatactct tcctttttca atattattga agcatttatc agggttattg
     3361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3421 cacgaattgg ccagcgctgc catttttggg gtgaggccgt tcgcggccga ggggcgcagc
     3481 ccctgggggg atgggaggcc cgcgttagcg ggccgggagg gttcgagaag ggggggcacc
     3541 ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa caaggtttat
     3601 aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa acgggcggaa
     3661 acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca ataggtgcgc
     3721 ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc atctgtcagt
     3781 agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg tccacatcat
     3841 ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg gccagctcca
     3901 cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg gtgagtcggc
     3961 ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt ccgcgaggta
     4021 tccacaacgc cggcggccgc ggtgtctcgc acacggcttc gacggcgttt ctggcgcgtt
     4081 tgcagggcca tagacggccg ccagcccagc ggcgagggca accagcccgg tgagcgtcgc
     4141 aaaggagatc ctgatctgac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc
     4201 tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacaaattg
     4261 acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg tggatgcagt
     4321 gggccc
//
