LOCUS       40924_13135        5137 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5243, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5137)
  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   2  (bases 1 to 5137)
  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   3  (bases 1 to 5137)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5137)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "2007"; volume: "189"; issue: "22"; pages: 
            "8059-8072"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-JUL-2007) Plant Pathology, Cornell University, 334 Plant 
            Science, Ithaca, NY 14853, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5137
                     /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             complement(348..1136)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             1425..3233
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     protein_bind    complement(3314..3347)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(3377..3396)
                     /label=priming site 2
                     /note="priming site 2"
     rep_origin      complement(3396..3779)
                     /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(3852..4709)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4710..4814)
                     /label=AmpR promoter
     terminator      4905..4991
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5083..5110
                     /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 atctttattt gccgactacc
      361 ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc
      421 aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac
      481 tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg
      541 gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc
      601 cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt
      661 tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct
      721 cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct
      781 gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc
      841 cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc
      901 tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca
      961 tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca
     1021 tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg
     1081 acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcatgatg
     1141 tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct ccataacatc
     1201 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtacc
     1261 ccaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt
     1321 tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta cagcttacga
     1381 accgaacagg cttatgtcca gatccagcta aggaggagtc gaccatggtc cgtcctgtag
     1441 aaaccccaac ccgtgaaatc aaaaaactcg acggcctgtg ggcattcagt ctggatcgcg
     1501 aaaactgtgg aattgatcag cgttggtggg aaagcgcgtt acaagaaagc cgggcaattg
     1561 ctgtgccagg cagttttaac gatcagttcg ccgatgcaga tattcgtaat tatgcgggca
     1621 acgtctggta tcagcgcgaa gtctttatac cgaaaggttg ggcaggccag cgtatcgtgc
     1681 tgcgtttcga tgcggtcact cattacggca aagtgtgggt caataatcag gaagtgatgg
     1741 agcatcaggg cggctatacg ccatttgaag ccgatgtcac gccgtatgtt attgccggga
     1801 aaagtgtacg tatcaccgtt tgtgtgaaca acgaactgaa ctggcagact atcccgccgg
     1861 gaatggtgat taccgacgaa aacggcaaga aaaagcagtc ttacttccat gatttcttta
     1921 actatgccgg aatccatcgc agcgtaatgc tctacaccac gccgaacacc tgggtggacg
     1981 atatcaccgt ggtgacgcat gtcgcgcaag actgtaacca cgcgtctgtt gactggcagg
     2041 tggtggccaa tggtgatgtc agcgttgaac tgcgtgatgc ggatcaacag gtggttgcaa
     2101 ctggacaagg cactagcggg actttgcaag tggtgaatcc gcacctctgg caaccgggtg
     2161 aaggttatct ctatgaactg tgcgtcacag ccaaaagcca gacagagtgt gatatctacc
     2221 cgcttcgcgt cggcatccgg tcagtggcag tgaagggcca acagttcctg attaaccaca
     2281 aaccgttcta ctttactggc tttggtcgtc atgaagatgc ggacttacgt ggcaaaggat
     2341 tcgataacgt gctgatggtg cacgaccacg cattaatgga ctggattggg gccaactcct
     2401 accgtacctc gcattaccct tacgctgaag agatgctcga ctgggcagat gaacatggca
     2461 tcgtggtgat tgatgaaact gctgctgtcg gctttaacct ctctttaggc attggtttcg
     2521 aagcgggcaa caagccgaaa gaactgtaca gcgaagaggc agtcaacggg gaaactcagc
     2581 aagcgcactt acaggcgatt aaagagctga tagcgcgtga caaaaaccac ccaagcgtgg
     2641 tgatgtggag tattgccaac gaaccggata cccgtccgca aggtgcacgg gaatatttcg
     2701 cgccactggc ggaagcaacg cgtaaactcg acccgacgcg tccgatcacc tgcgtcaatg
     2761 taatgttctg cgacgctcac accgatacca tcagcgatct ctttgatgtg ctgtgcctga
     2821 accgttatta cggatggtat gtccaaagcg gcgatttgga aacggcagag aaggtactgg
     2881 aaaaagaact tctggcctgg caggagaaac tgcatcagcc gattatcatc accgaatacg
     2941 gcgtggatac gttagccggg ctgcactcaa tgtacaccga catgtggagt gaagagtatc
     3001 agtgtgcatg gctggatatg tatcaccgcg tctttgatcg cgtcagcgcc gtcgtcggtg
     3061 aacaggtatg gaatttcgcc gattttgcga cctcgcaagg catattgcgc gttggcggta
     3121 acaagaaagg gatcttcact cgcgaccgca aaccgaagtc ggcggctttt ctgctgcaaa
     3181 aacgctggac tggcatgaac ttcggtgaaa aaccgcagca gggaggcaaa caatgaatca
     3241 acaactctcc tggcgcacca tcgtcggcta cagcctcggt ggggaataga tcccccagct
     3301 tcaaaagcgc tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaacta
     3361 aggaggatat tcatatggac catggctaat tcccatgtca gccgttaagt gttcctgtgt
     3421 cactgaaaat tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt
     3481 tgtccacaac cgttaaacct taaaagcttt aaaagcctta tatattcttt tttttcttat
     3541 aaaacttaaa accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa
     3601 catgagagct tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac
     3661 gttagccatg agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct
     3721 tagtacgtga aacatgagag cttagtacgt actatcaaca ggttgaactg cggatcttgc
     3781 ggccgcaaaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3841 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3901 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3961 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4021 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4081 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4141 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4201 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4261 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4321 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4381 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4441 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4501 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4561 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4621 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4681 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4741 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4801 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg catcgatggc cccccgatgg
     4861 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     4921 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     4981 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     5041 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     5101 atggcctttt tgcgtggcca gtgccaagct tgcatgc
//
