LOCUS       sequence-405613-        2836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing an origin of transfer sequence from 
            pPtPBR11 required for making plasmids mobilizable for conjugation 
            with CE overhangs used to build level 1 constructs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2836)
  AUTHORS   Pollak B, Matute T, Nunez I, Cerda A, Lopez C, Vargas V, Kan A, 
            Bielinski V, von Dassow P, Dupont CL, Federici F
  TITLE     Universal loop assembly: open, efficient and cross-kingdom DNA 
            fabrication.
  JOURNAL   Synth Biol (Oxf). 2020;5(1):ysaa001. doi: 10.1093/synbio/ysaa001. 
            Epub 2020 Feb 5.
  PUBMED    32161816
REFERENCE   2  (bases 1 to 2836)
  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: "Synth Biol 
            (Oxf)."; date: "2020"; volume: "5(1):ysaa001. doi"; pages: " 
            10.1093/synbio/ysaa001. Epub 2020 Feb 5"
COMMENT     Sequence Label: pL0_CE-OriT
FEATURES             Location/Qualifiers
     source          1..2836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1199..1570)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELVRINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1603..1712
                     /note="incP origin of transfer"
     rep_origin      complement(1990..2578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2070..2089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2669..2836,1..624))
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
       61 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
      121 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
      181 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
      241 gtcgatcgtg gctggctcga agataccagc aagaatgtca ttgcgctgcc attctccaaa
      301 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
      361 gacttctaca gcgcggagaa tctcactctc tccaggggaa gccgaagttt ccaaaaggtc
      421 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
      481 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
      541 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
      601 ttcggcgatc accgcttccc tcatgcgaaa cgatcctcat cctgtctctt gatcagatat
      661 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
      721 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctaagcttgc
      781 atgcctgcaa gtcgactcta gaggagcggt gatcacaggc agcaacgctc tgtcatcgtt
      841 acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca gcttagttgc
      901 cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa cggctctccc
      961 gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctc
     1021 attactcgca tccattctca ggctgtctcg tctcgtctca ggtctcaaat ggatcgtctt
     1081 gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgcccttct tgatggagcg
     1141 catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc
     1201 atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct
     1261 cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt
     1321 cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcgga
     1381 ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg
     1441 ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca atcgcccttc
     1501 gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt
     1561 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     1621 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     1681 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     1741 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     1801 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaagatgct ttgagaccac
     1861 caggatacat agattaccac aactccgagc ccttccaccc acagaatcag gggataacgc
     1921 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1981 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2041 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2101 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2161 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2221 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2281 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2341 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2401 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2461 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2521 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2581 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2641 gattttggtc atgcattcta ggtgattatt atttgccgac taccttggtg atctcgcctt
     2701 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt
     2761 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct
     2821 ccattgccca gtcggc
//
