LOCUS       40924_33407        4147 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pNML54, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4147)
  AUTHORS   Rozwadowski K, Yang W, Kagale S.
  TITLE     Homologous recombination-mediated cloning and manipulation of 
            genomic DNA regions using Gateway and recombineering systems
  JOURNAL   BMC Biotechnol. 8, 88 (2008)
  PUBMED    19014699
REFERENCE   2  (bases 1 to 4147)
  AUTHORS   Rozwadowski KL, Yang W, Kagale S.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-OCT-2008) Molecular Genetics, Agriculture and 
            Agri-Food Canada, 107 Science Place, Saskatoon, Saskatchewan S7N 
            0X2, Canada
REFERENCE   3  (bases 1 to 4147)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4147)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol. 8, 88 (2008)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-OCT-2008) Molecular Genetics, Agriculture and Agri-Food Canada, 
            107 Science Place, Saskatoon, Saskatchewan S7N 0X2, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(106..130)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    171..278
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     misc_feature    complement(342..366)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      444..1075
                     /label=oriV
                     /note="incP origin of replication"
     CDS             1918..2592
                     /codon_start=1
                     /gene="aad9"
                     /product="Aad9"
                     /label=aad9
                     /note="spectinomycin resistance"
                     /protein_id="ACL35309.1"
                     /translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
                     GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
                     QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
                     MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
                     KL"
     gene            1918..2592
                     /gene="aad9"
                     /label=aad9
     CDS             2953..4098
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
ORIGIN
        1 tgagatctcc tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc
       61 ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta cccgccaata
      121 tatcctgtca aacactgata gtttaaactg aaggcgggaa acgacaatct gagctccacc
      181 gcggtggcgg ccgctctaga actagtggat cccccgggct gcaggaattc gatatcaagc
      241 ttatcgatac cgtcgacctc gagggggggc ccggtaccaa aaccacccca gtacattaaa
      301 aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc
      361 ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat
      421 acaggcagcc catcagtcca ctagacgctc accgggctgg ttgccctcgc cgctgggctg
      481 gcggccgtct atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac
      541 cgcggccgcc ggcgttgtgg atacctcgcg gaaaacttgg ccctcactga cagatgaggg
      601 gcggacgttg acacttgagg ggccgactca cccggcgcgg cgttgacaga tgaggggcag
      661 gctcgatttc ggccggcgac gtggagctgg ccagcctcgc aaatcggcga aaacgcctga
      721 ttttacgcga gtttcccaca gatgatgtgg acaagcctgg ggataagtgc cctgcggtat
      781 tgacacttga ggggcgcgac tactgacaga tgaggggcgc gatccttgac acttgagggg
      841 cagagtgctg acagatgagg ggcgcaccta ttgacatttg aggggctgtc cacaggcaga
      901 aaatccagca tttgcaaggg tttccgcccg tttttcggcc accgctaacc tgtcttttaa
      961 cctgctttta aaccaatatt tataaacctt gtttttaacc agggctgcgc cctgtgcgcg
     1021 tgaccgcgca cgccgaaggg gggtgccccc ccttctcgaa ccctcccggc ccgctctcga
     1081 gttggcagca tcacccataa ttgtggtttc aaaatcggct ccgtcgatac tatgttatac
     1141 gccaactttg aaaacaactt tgaaaaagct gttttctggt atttaaggtt ttagaatgca
     1201 aggaacagtg aattggagtt cgtcttgtta taattagctt cttggggtat ctttaaatac
     1261 tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag aatatcaccg gaattgaaaa
     1321 aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct gctaaggtat
     1381 ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc cggtataaag
     1441 ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa ggaaagctgc
     1501 ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg ctcatgagct
     1561 agccgttcgt gaatacatgt tataataact ataactaata acgtaacgtg actggcaaga
     1621 gatattttta aaacaatgaa taggtttaca cttactttag ttttatggaa atgaaagatc
     1681 atatcatata taatctagaa taaaattaac taaaataatt attatctaga taaaaaattt
     1741 agaagccaat gaaatctata aataaactaa attaagttta tttaattaac aactatggat
     1801 ataaaatagg tactaatcaa aatagtgagg aggatatatt tgaatacata cgaacaaatt
     1861 aataaagtga aaaaaatact tcggaaacat ttaaaaaata accttattgg tacttacatg
     1921 tttggatcag gagttgagag tggactaaaa ccaaatagtg atcttgactt tttagtcgtc
     1981 gtatctgaac cattgacaga tcaaagtaaa gaaatactta tacaaaaaat tagacctatt
     2041 tcaaaaaaaa taggagataa aagcaactta cgatatattg aattaacaat tattattcag
     2101 caagaaatgg taccgtggaa tcatcctccc aaacaagaat ttatttatgg agaatggtta
     2161 caagagcttt atgaacaagg atacattcct cagaaggaat taaattcaga tttaaccata
     2221 atgctttacc aagcaaaacg aaaaaataaa agaatatacg gaaattatga cttagaggaa
     2281 ttactacctg atattccatt ttctgatgtg agaagagcca ttatggattc gtcagaggaa
     2341 ttaatagata attatcagga tgatgaaacc aactctatat taactttatg ccgtatgatt
     2401 ttaactatgg acacgggtaa aatcatacca aaagatattg cgggaaatgc agtggctgaa
     2461 tcttctccat tagaacatag ggagagaatt ttgttagcag ttcgtagtta tcttggagag
     2521 aatattgaat ggactaatga aaatgtaaat ttaactataa actatttaaa taacagatta
     2581 aaaaaattat aaaaaaattg aaaaaatggt ggaaacactt ttgttattat attttactgg
     2641 atgaattgtt ttagtaccta gatgtggcgc aacgatgccg gcgacaagca ggagcgcacc
     2701 gacttcttcc gcatcaagtg ttttggctct caggccgagg cccacggcaa gtatttgggc
     2761 aaggggtcgc tggtattcgt gcagggcaag attcggaata ccaagtacga gaaggacggc
     2821 cagacggtct acgggaccga cttcattgcc gataaggtgg attatctgga caccaaggca
     2881 ccaggcgggt caaatcagga ataagggcac attgccccgg cgtgagtcgg ggcaatcccg
     2941 caaggagggt gaatgaatcg gacgtttgac cggaaggcat acaggcaaga actgatcgac
     3001 gcggggtttt ccgccgagga tgccgaaacc atcgcaagcc gcaccgtcat gcgtgcgccc
     3061 cgcgaaacct tccagtccgt cggctcgatg gtccagcaag ctacggccaa gatcgagcgc
     3121 gacagcgtgc aactggctcc ccctgccctg cccgcgccat cggccgccgt ggagcgttcg
     3181 cgtcgtctcg aacaggaggc ggcaggtttg gcgaagtcga tgaccatcga cacgcgagga
     3241 actatgacga ccaagaagcg aaaaaccgcc ggcgaggacc tggcaaaaca ggtcagcgag
     3301 gccaagcagg ccgcgttgct gaaacacacg aagcagcaga tcaaggaaat gcagctttcc
     3361 ttgttcgata ttgcgccgtg gccggacacg atgcgagcga tgccaaacga cacggcccgc
     3421 tctgccctgt tcaccacgcg caacaagaaa atcccgcgcg aggcgctgca aaacaaggtc
     3481 attttccacg tcaacaagga cgtgaagatc acctacaccg gcgtcgagct gcgggccgac
     3541 gatgacgaac tggtgtggca gcaggtgttg gagtacgcga agcgcacccc tatcggcgag
     3601 ccgatcacct tcacgttcta cgagctttgc caggacctgg gctggtcgat caatggccgg
     3661 tattacacga aggccgagga atgcctgtcg cgcctacagg cgacggcgat gggcttcacg
     3721 tccgaccgcg ttgggcacct ggaatcggtg tcgctgctgc accgcttccg cgtcctggac
     3781 cgtggcaaga aaacgtcccg ttgccaggtc ctgatcgacg aggaaatcgt cgtgctgttt
     3841 gctggcgacc actacacgaa attcatatgg gagaagtacc gcaagctgtc gccgacggcc
     3901 cgacggatgt tcgactattt cagctcgcac cgggagccgt acccgctcaa gctggaaacc
     3961 ttccgcctca tgtgcggatc ggattccacc cgcgtgaaga agtggcgcga gcaggtcggc
     4021 gaagcctgcg aagagttgcg aggcagcggc ctggtggaac acgcctgggt caatgatgac
     4081 ctggtgcatt gcaaacgcta gggccttgtg gggtcagttc cggctggggg ttcagccagc
     4141 gctttac
//
