LOCUS       sequence-465757-        4585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing the Insert 4 to be assembled in POC1355.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4585)
  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 4585)
  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: POC1346
FEATURES             Location/Qualifiers
     source          1..4585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    357..381
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(537..1685)
                     /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(2021..2609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2101..2120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2780..3574)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(3575..3679)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3766..4397
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4489..4513
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cactctgtga cttggtctca ttacgtactg tattcgaccc attcggtgat gtcatatggc
       61 gtggtgccca gacgacgcta gaccgtagac tatggtcatt gtgggaagga atcgcgactt
      121 gcggttctca cggttgagag tggaccggcc gagctgacat tctaacgagt cgttgccact
      181 tctttgctta ttacttcata aactccaaat tcggcatgat cggagtcggc gagcagtcag
      241 taccggggcg atgtgggtta aggacgccaa gttttccatg cagaacacgt ggcggtcatg
      301 acgtcgatcg ggtacagatg agaccaagtc acgaagtggt ttaaactatc agtgtttgac
      361 aggatatatt ggcgggtaaa cctaagagaa aagagcgttt attagaataa tcggatattt
      421 aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt
      481 tccccagatc aggcgctggc tgctgaaccc ccagccggaa ctgaccccac aaggccctag
      541 cgtttgcaat gcaccaggtc atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc
      601 aactcttcgc aggcttcgcc gacctgctcg cgccacttct tcacgcgggt ggaatccgat
      661 ccgcacatga ggcggaaggt ttccagcttg agcgggtacg gctcccggtg cgagctgaaa
      721 tagtcgaaca tccgtcgggc cgtcggcgac agcttgcggt acttctccca tatgaatttc
      781 gtgtagtggt cgccagcaaa cagcacgacg atttcctcgt cgatcaggac ctggcaacgg
      841 gacgttttct tgccacggtc caggacgcgg aagcggtgca gcagcgacac cgattccagg
      901 tgcccaacgc ggtcggacgt gaagcccatc gccgtcgcct gtaggcgcga caggcattcc
      961 tcggccttcg tgtaataccg gccattgatc gaccagccca ggtcctggca aagctcgtag
     1021 aacgtgaagg tgatcggctc gccgataggg gtgcgcttcg cgtactccaa cacctgctgc
     1081 cacaccagtt cgtcatcgtc ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc
     1141 ttgttgacgt ggaaaatgac cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc
     1201 gtggtgaaca gggcagagcg ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac
     1261 ggcgcaatat cgaacaagga aagctgcatt tccttgatct gctgcttcgt gtgtttcagc
     1321 aacgcggcct gcttggcctc gctgacctgt tttgccaggt cctcgccggc ggtttttcgc
     1381 ttcttggtcg tcatagttcc tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc
     1441 tcctgttcaa gacgacgcga acgctccacg gcggccgatg gcgcgggcag ggcaggggga
     1501 gccagttgca cgctgtcgcg ctcgatcttg gccgtagctt gctggaccat cgagccgacg
     1561 gactggaagg tttcgcgggg cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc
     1621 tcggcggaaa accccgcgtc gatcagttct tgcctgtatg ccttccggtc aaacgtccga
     1681 ttcattcacc ctccttgcgg gattgccccg actcacgccg gggcaatgtg cccttattcc
     1741 tgatttgacc cgcctggtgc cttggtgtcc agataatcca ccttatcggc aatgaagtcg
     1801 gtcccgtaga ccgtctggcc gtccttctcg tacttggtat tccgaatctt gccctgcacg
     1861 aataccagcg accccttgcc caaatacttg ccgtgggcct cggcctgaga gccaaaacac
     1921 ttgatgcgga agaagtcggt gcgctcctgc ttgtcgccgg catcgttgcg ccacatctag
     1981 gatctgccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2041 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2101 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2161 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2221 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2281 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2341 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2401 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2461 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2521 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2581 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2641 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2701 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2761 agtaaacttg gtctgacagc taaaacaatt catccagtaa aatataatat tttattttct
     2821 cccaatcagg cttgatcccc agtaagtcaa aaaatagctc gacatactgt tcttccccga
     2881 tatcctccct gagcgaccgg acgcagaagg caatgtcata ccacttgtcc gccctgccgc
     2941 ttctcccaag atcaataaag ccacttactt tgccatcttt cacaaagatg ttgctgtctc
     3001 ccaggtcgcc gtgggaaaag acaagttcct cttcgggctt ttccgtcttt aaaaaatcat
     3061 acagctcgcg cggatcttta aatggagtgt cctcttccca gttttcgcaa tccacatcgg
     3121 ccagatcgtt attcagtaag taatccaatt cggctaagcg gctgtctaag ctattcgtat
     3181 agggacaatc cgatatgtcg atggagtgaa agagcctgat gcactccgca tacagctcga
     3241 taatcttttc agggctttgt tcatcttcat actcttccga gcaaaggacg ccatcggcct
     3301 cactcatgag cagattgctc cagccatcat gccgttcaaa gtgcaggacc tttggaacag
     3361 gcagctttcc ttccagccat agcatcatgt ccttttcccg ttccacatca taggtggtcc
     3421 ctttataccg gctgtccgtc atttttaaat ataggttttc attttctccc accagcttat
     3481 ataccttagc aggagacatt ccttccgtat cttttacgca gcggtatttt tcgatcagtt
     3541 ttttcaattc cggtgatatt ctcattttag ccatactctt cctttttcaa tattattgaa
     3601 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3661 aacaaatagg ggttccgcgc acgaattggc cagcgctgcc atttttgggg tgaggccgtt
     3721 cgcggccgag gggcgcagcc cctgggggga tgggaggccc gcgttagcgg gccgggaggg
     3781 ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca cgcgcacagg gcgcagccct
     3841 ggttaaaaac aaggtttata aatattggtt taaaagcagg ttaaaagaca ggttagcggt
     3901 ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt tctgcctgtg gacagcccct
     3961 caaatgtcaa taggtgcgcc cctcatctgt cagcactctg cccctcaagt gtcaaggatc
     4021 gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca ataccgcagg gcacttatcc
     4081 ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa tcaggcgttt tcgccgattt
     4141 gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc ctgcccctca tctgtcaacg
     4201 ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc ccctcatctg tcagtgaggg
     4261 ccaagttttc cgcgaggtat ccacaacgcc ggcggccgcg gtgtctcgca cacggcttcg
     4321 acggcgtttc tggcgcgttt gcagggccat agacggccgc cagcccagcg gcgagggcaa
     4381 ccagcccggt gagcgtcgca aaggagatcc tgatctgact gatgggctgc ctgtatcgag
     4441 tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata
     4501 ttgtggtgta aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat
     4561 gtactggggt ggatgcagtg ggccc
//
