LOCUS       40924_13130        3249 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5242, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3249)
  AUTHORS   Wei CF, Kvitko BH, Shimizu R, Crabill E, Alfano JR, Lin NC, Martin 
            GB, Huang HC, Collmer A.
  TITLE     A Pseudomonas syringae pv. tomato DC3000 mutant lacking the type III
            effector HopQ1-1 is able to cause disease in the model plant 
            Nicotiana benthamiana
  JOURNAL   Plant J. 51 (1), 32-46 (2007)
  PUBMED    17559511
REFERENCE   2  (bases 1 to 3249)
  AUTHORS   Kvitko BH, Ramos AR, Morello JE, Oh HS, Collmer A.
  TITLE     Identification of harpins in Pseudomonas syringae pv. tomato DC3000,
            which are functionally similar to HrpK1 in promoting translocation 
            of type III secretion system effectors
  JOURNAL   J. Bacteriol. 189 (22), 8059-8072 (2007)
  PUBMED    17873033
REFERENCE   3  (bases 1 to 3249)
  AUTHORS   Kvitko BH, Ramos AR, Morello JE, Oh H-S., Collmer A.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUL-2007) Plant Pathology, Cornell University, 334 
            Plant Science, Ithaca, NY 14853, USA
REFERENCE   4  (bases 1 to 3249)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3249)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant J."; 
            date: "2007"; volume: "51"; issue: "1"; pages: "32-46"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "2007"; volume: "189"; issue: "22"; pages: 
            "8059-8072"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (10-JUL-2007) Plant Pathology, Cornell University, 334 Plant 
            Science, Ithaca, NY 14853, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=priming site 1
                     /note="priming site 1"
     misc_recomb     51..98
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(51..98)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             608..1396
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     protein_bind    complement(1426..1459)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(1489..1508)
                     /label=priming site 2
                     /note="priming site 2"
     rep_origin      complement(1508..1891)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1964..2821)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2822..2926)
                     /label=AmpR promoter
     terminator      3017..3103
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3195..3222
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 agattgcagc attacacgtc ttgagcgatt gtgtaggctg gagctgcttc gaagttccta
       61 tactttctag agaataggaa cttcggaata ggaacttcaa gatcccctca cgctgccgca
      121 agcactcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga
      181 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
      241 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
      301 ttttatggac agcaagcgaa ccggaattgc cagctgggag atctggacat aagcctgttc
      361 ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
      421 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
      481 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
      541 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
      601 aaacatcatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
      661 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
      721 ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct
      781 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
      841 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
      901 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
      961 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     1021 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     1081 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     1141 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     1201 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     1261 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     1321 cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac
     1381 caaggtagtc ggcaaataaa gatcccccag cttcaaaagc gctctgaagt tcctatactt
     1441 tctagagaat aggaacttcg gaataggaac taaggaggat attcatatgg accatggcta
     1501 attcccatgt cagccgttaa gtgttcctgt gtcactgaaa attgctttga gaggctctaa
     1561 gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaagct
     1621 ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa
     1681 gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag
     1741 agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa
     1801 catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac
     1861 gtactatcaa caggttgaac tgcggatctt gcggccgcaa aaattaaaaa tgaagtttta
     1921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2941 aagtgccacc tgcatcgatg gccccccgat ggtagtgtgg ggtctcccca tgcgagagta
     3001 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     3061 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     3121 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     3181 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtggc cagtgccaag
     3241 cttgcatgc
//
