LOCUS       40924_13145        5155 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5250, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5155)
  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 5155)
  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 5155)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5155)
  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..5155
                     /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             448..2256
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     CDS             2347..3138
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(3332..3365)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(3395..3414)
                     /label=priming site 2
                     /note="priming site 2"
     rep_origin      complement(3414..3797)
                     /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(3870..4727)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4728..4832)
                     /label=AmpR promoter
     terminator      4923..5009
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5101..5128
                     /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 cagctggggc gccctctggt aaggttggga
      361 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
      421 caagatccag ctaaggagga gtcgaccatg gtccgtcctg tagaaacccc aacccgtgaa
      481 atcaaaaaac tcgacggcct gtgggcattc agtctggatc gcgaaaactg tggaattgat
      541 cagcgttggt gggaaagcgc gttacaagaa agccgggcaa ttgctgtgcc aggcagtttt
      601 aacgatcagt tcgccgatgc agatattcgt aattatgcgg gcaacgtctg gtatcagcgc
      661 gaagtcttta taccgaaagg ttgggcaggc cagcgtatcg tgctgcgttt cgatgcggtc
      721 actcattacg gcaaagtgtg ggtcaataat caggaagtga tggagcatca gggcggctat
      781 acgccatttg aagccgatgt cacgccgtat gttattgccg ggaaaagtgt acgtatcacc
      841 gtttgtgtga acaacgaact gaactggcag actatcccgc cgggaatggt gattaccgac
      901 gaaaacggca agaaaaagca gtcttacttc catgatttct ttaactatgc cggaatccat
      961 cgcagcgtaa tgctctacac cacgccgaac acctgggtgg acgatatcac cgtggtgacg
     1021 catgtcgcgc aagactgtaa ccacgcgtct gttgactggc aggtggtggc caatggtgat
     1081 gtcagcgttg aactgcgtga tgcggatcaa caggtggttg caactggaca aggcactagc
     1141 gggactttgc aagtggtgaa tccgcacctc tggcaaccgg gtgaaggtta tctctatgaa
     1201 ctgtgcgtca cagccaaaag ccagacagag tgtgatatct acccgcttcg cgtcggcatc
     1261 cggtcagtgg cagtgaaggg ccaacagttc ctgattaacc acaaaccgtt ctactttact
     1321 ggctttggtc gtcatgaaga tgcggactta cgtggcaaag gattcgataa cgtgctgatg
     1381 gtgcacgacc acgcattaat ggactggatt ggggccaact cctaccgtac ctcgcattac
     1441 ccttacgctg aagagatgct cgactgggca gatgaacatg gcatcgtggt gattgatgaa
     1501 actgctgctg tcggctttaa cctctcttta ggcattggtt tcgaagcggg caacaagccg
     1561 aaagaactgt acagcgaaga ggcagtcaac ggggaaactc agcaagcgca cttacaggcg
     1621 attaaagagc tgatagcgcg tgacaaaaac cacccaagcg tggtgatgtg gagtattgcc
     1681 aacgaaccgg atacccgtcc gcaaggtgca cgggaatatt tcgcgccact ggcggaagca
     1741 acgcgtaaac tcgacccgac gcgtccgatc acctgcgtca atgtaatgtt ctgcgacgct
     1801 cacaccgata ccatcagcga tctctttgat gtgctgtgcc tgaaccgtta ttacggatgg
     1861 tatgtccaaa gcggcgattt ggaaacggca gagaaggtac tggaaaaaga acttctggcc
     1921 tggcaggaga aactgcatca gccgattatc atcaccgaat acggcgtgga tacgttagcc
     1981 gggctgcact caatgtacac cgacatgtgg agtgaagagt atcagtgtgc atggctggat
     2041 atgtatcacc gcgtctttga tcgcgtcagc gccgtcgtcg gtgaacaggt atggaatttc
     2101 gccgattttg cgacctcgca aggcatattg cgcgttggcg gtaacaagaa agggatcttc
     2161 actcgcgacc gcaaaccgaa gtcggcggct tttctgctgc aaaaacgctg gactggcatg
     2221 aacttcggtg aaaaaccgca gcagggaggc aaacaatgaa tcaacaactc tcctggcgca
     2281 ccatcgtcgg ctacagcctc ggtggggaat ggatctgatc aagagacagg atgaggatcg
     2341 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2401 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2461 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2521 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2581 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2641 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2701 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2761 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     2821 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     2881 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2941 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3001 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3061 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3121 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3181 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3241 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3301 tcttcgccca ccccagcttc aaaagcgctc tgaagttcct atactttcta gagaatagga
     3361 acttcggaat aggaactaag gaggatattc atatggacca tggctaattc ccatgtcagc
     3421 cgttaagtgt tcctgtgtca ctgaaaattg ctttgagagg ctctaagggc ttctcagtgc
     3481 gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa aagccttata
     3541 tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg ctgatttata
     3601 ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtta
     3661 gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg agagcttagt
     3721 acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac tatcaacagg
     3781 ttgaactgcg gatcttgcgg ccgcaaaaat taaaaatgaa gttttaaatc aatctaaagt
     3841 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3901 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3961 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4021 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4081 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4141 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4201 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4261 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4321 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4381 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4441 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4501 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4561 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4621 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4681 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4741 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4801 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgca
     4861 tcgatggccc cccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     4921 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     4981 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     5041 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     5101 agaaggccat cctgacggat ggcctttttg cgtggccagt gccaagcttg catgc
//
