LOCUS       40924_13125        4833 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5212, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4833)
  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 4833)
  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 4833)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4833)
  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..4833
                     /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)."
     promoter        345..373
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             562..1092
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             1121..2929
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     protein_bind    complement(3010..3043)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(3073..3092)
                     /label=priming site 2
                     /note="priming site 2"
     rep_origin      complement(3092..3475)
                     /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(3548..4405)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4406..4510)
                     /label=AmpR promoter
     terminator      4601..4687
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4779..4806
                     /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 atctttgaca taagcctgtt
      361 cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
      421 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
      481 ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
      541 tgatgttatg gagcagcaac gatgttacgc agcagcaacg atgttacgca gcagggcagt
      601 cgccctaaaa caaagttagg tggctcaagt atgggcatca ttcgcacatg taggctcggc
      661 cctgaccaag tcaaatccat gcgggctgct cttgatcttt tcggtcgtga gttcggagac
      721 gtagccacct actcccaaca tcagccggac tccgattacc tcgggaactt gctccgtagt
      781 aagacattca tcgcgcttgc tgccttcgac caagaagcgg ttgttggcgc tctcgcggct
      841 tacgttctgc ccaggtttga gcagccgcgt agtgagatct atatctatga tctcgcagtc
      901 tccggcgagc accggaggca gggcattgcc accgcgctca tcaatctcct caagcatgag
      961 gccaacgcgc ttggtgctta tgtgatctac gtgcaagcag attacggtga cgatcccgca
     1021 gtggctctct atacaaagtt gggcatacgg gaagaagtga tgcactttga tatcgaccca
     1081 agtaccgcca cctaaggatc cagctaagga ggagtcgacc atggtccgtc ctgtagaaac
     1141 cccaacccgt gaaatcaaaa aactcgacgg cctgtgggca ttcagtctgg atcgcgaaaa
     1201 ctgtggaatt gatcagcgtt ggtgggaaag cgcgttacaa gaaagccggg caattgctgt
     1261 gccaggcagt tttaacgatc agttcgccga tgcagatatt cgtaattatg cgggcaacgt
     1321 ctggtatcag cgcgaagtct ttataccgaa aggttgggca ggccagcgta tcgtgctgcg
     1381 tttcgatgcg gtcactcatt acggcaaagt gtgggtcaat aatcaggaag tgatggagca
     1441 tcagggcggc tatacgccat ttgaagccga tgtcacgccg tatgttattg ccgggaaaag
     1501 tgtacgtatc accgtttgtg tgaacaacga actgaactgg cagactatcc cgccgggaat
     1561 ggtgattacc gacgaaaacg gcaagaaaaa gcagtcttac ttccatgatt tctttaacta
     1621 tgccggaatc catcgcagcg taatgctcta caccacgccg aacacctggg tggacgatat
     1681 caccgtggtg acgcatgtcg cgcaagactg taaccacgcg tctgttgact ggcaggtggt
     1741 ggccaatggt gatgtcagcg ttgaactgcg tgatgcggat caacaggtgg ttgcaactgg
     1801 acaaggcact agcgggactt tgcaagtggt gaatccgcac ctctggcaac cgggtgaagg
     1861 ttatctctat gaactgtgcg tcacagccaa aagccagaca gagtgtgata tctacccgct
     1921 tcgcgtcggc atccggtcag tggcagtgaa gggccaacag ttcctgatta accacaaacc
     1981 gttctacttt actggctttg gtcgtcatga agatgcggac ttacgtggca aaggattcga
     2041 taacgtgctg atggtgcacg accacgcatt aatggactgg attggggcca actcctaccg
     2101 tacctcgcat tacccttacg ctgaagagat gctcgactgg gcagatgaac atggcatcgt
     2161 ggtgattgat gaaactgctg ctgtcggctt taacctctct ttaggcattg gtttcgaagc
     2221 gggcaacaag ccgaaagaac tgtacagcga agaggcagtc aacggggaaa ctcagcaagc
     2281 gcacttacag gcgattaaag agctgatagc gcgtgacaaa aaccacccaa gcgtggtgat
     2341 gtggagtatt gccaacgaac cggatacccg tccgcaaggt gcacgggaat atttcgcgcc
     2401 actggcggaa gcaacgcgta aactcgaccc gacgcgtccg atcacctgcg tcaatgtaat
     2461 gttctgcgac gctcacaccg ataccatcag cgatctcttt gatgtgctgt gcctgaaccg
     2521 ttattacgga tggtatgtcc aaagcggcga tttggaaacg gcagagaagg tactggaaaa
     2581 agaacttctg gcctggcagg agaaactgca tcagccgatt atcatcaccg aatacggcgt
     2641 ggatacgtta gccgggctgc actcaatgta caccgacatg tggagtgaag agtatcagtg
     2701 tgcatggctg gatatgtatc accgcgtctt tgatcgcgtc agcgccgtcg tcggtgaaca
     2761 ggtatggaat ttcgccgatt ttgcgacctc gcaaggcata ttgcgcgttg gcggtaacaa
     2821 gaaagggatc ttcactcgcg accgcaaacc gaagtcggcg gcttttctgc tgcaaaaacg
     2881 ctggactggc atgaacttcg gtgaaaaacc gcagcaggga ggcaaacaat gaatcaacaa
     2941 ctctcctggc gcaccatcgt cggctacagc ctcggtgggg aatagatccc ccagcttcaa
     3001 aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag gaactaagga
     3061 ggatattcat atggaccatg gctaattccc atgtcagccg ttaagtgttc ctgtgtcact
     3121 gaaaattgct ttgagaggct ctaagggctt ctcagtgcgt tacatccctg gcttgttgtc
     3181 cacaaccgtt aaaccttaaa agctttaaaa gccttatata ttcttttttt tcttataaaa
     3241 cttaaaacct tagaggctat ttaagttgct gatttatatt aattttattg ttcaaacatg
     3301 agagcttagt acgtgaaaca tgagagctta gtacgttagc catgagagct tagtacgtta
     3361 gccatgaggg tttagttcgt taaacatgag agcttagtac gttaaacatg agagcttagt
     3421 acgtgaaaca tgagagctta gtacgtacta tcaacaggtt gaactgcgga tcttgcggcc
     3481 gcaaaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3541 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3601 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3661 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3721 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3781 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3841 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3901 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3961 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4021 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4081 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4141 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4201 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4261 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4321 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4381 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4441 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4501 gggttccgcg cacatttccc cgaaaagtgc cacctgcatc gatggccccc cgatggtagt
     4561 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     4621 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     4681 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     4741 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     4801 cctttttgcg tggccagtgc caagcttgca tgc
//
