LOCUS       40924_13140        4699 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5249, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4699)
  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 4699)
  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 4699)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4699)
  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..4699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=priming site 1
                     /note="priming site 1"
     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)."
     misc_recomb     51..84
                     /label=FRT
                     /note="FRT"
     CDS             complement(134..772)
                     /codon_start=1
                     /gene="CmR"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /protein_id="ABS87324.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
     gene            complement(134..772)
                     /gene="CmR"
                     /label=CmR
     CDS             complement(845..2653)
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     promoter        complement(2680..2770)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2876..2909)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     primer_bind     complement(2939..2958)
                     /label=priming site 2
                     /note="priming site 2"
     rep_origin      complement(2958..3341)
                     /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(3414..4271)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4272..4376)
                     /label=AmpR promoter
     terminator      4467..4553
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4645..4672
                     /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 ggaacttcat ttaaatggcg cgccttacgc
      121 cccgccctgc cactcatcgc agtactgttg tattcattaa gcatctgccg acatggaagc
      181 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
      241 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
      301 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
      361 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
      421 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
      481 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
      541 tcattgccat acgtaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
      601 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
      661 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
      721 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
      781 ccttagatcc attccccacc gaggctgtag ccgacgatgg tgcgccagga gagttgttga
      841 ttcattgttt gcctccctgc tgcggttttt caccgaagtt catgccagtc cagcgttttt
      901 gcagcagaaa agccgccgac ttcggtttgc ggtcgcgagt gaagatccct ttcttgttac
      961 cgccaacgcg caatatgcct tgcgaggtcg caaaatcggc gaaattccat acctgttcac
     1021 cgacgacggc gctgacgcga tcaaagacgc ggtgatacat atccagccat gcacactgat
     1081 actcttcact ccacatgtcg gtgtacattg agtgcagccc ggctaacgta tccacgccgt
     1141 attcggtgat gataatcggc tgatgcagtt tctcctgcca ggccagaagt tctttttcca
     1201 gtaccttctc tgccgtttcc aaatcgccgc tttggacata ccatccgtaa taacggttca
     1261 ggcacagcac atcaaagaga tcgctgatgg tatcggtgtg agcgtcgcag aacattacat
     1321 tgacgcaggt gatcggacgc gtcgggtcga gtttacgcgt tgcttccgcc agtggcgcga
     1381 aatattcccg tgcaccttgc ggacgggtat ccggttcgtt ggcaatactc cacatcacca
     1441 cgcttgggtg gtttttgtca cgcgctatca gctctttaat cgcctgtaag tgcgcttgct
     1501 gagtttcccc gttgactgcc tcttcgctgt acagttcttt cggcttgttg cccgcttcga
     1561 aaccaatgcc taaagagagg ttaaagccga cagcagcagt ttcatcaatc accacgatgc
     1621 catgttcatc tgcccagtcg agcatctctt cagcgtaagg gtaatgcgag gtacggtagg
     1681 agttggcccc aatccagtcc attaatgcgt ggtcgtgcac catcagcacg ttatcgaatc
     1741 ctttgccacg taagtccgca tcttcatgac gaccaaagcc agtaaagtag aacggtttgt
     1801 ggttaatcag gaactgttgg cccttcactg ccactgaccg gatgccgacg cgaagcgggt
     1861 agatatcaca ctctgtctgg cttttggctg tgacgcacag ttcatagaga taaccttcac
     1921 ccggttgcca gaggtgcgga ttcaccactt gcaaagtccc gctagtgcct tgtccagttg
     1981 caaccacctg ttgatccgca tcacgcagtt caacgctgac atcaccattg gccaccacct
     2041 gccagtcaac agacgcgtgg ttacagtctt gcgcgacatg cgtcaccacg gtgatatcgt
     2101 ccacccaggt gttcggcgtg gtgtagagca ttacgctgcg atggattccg gcatagttaa
     2161 agaaatcatg gaagtaagac tgctttttct tgccgttttc gtcggtaatc accattcccg
     2221 gcgggatagt ctgccagttc agttcgttgt tcacacaaac ggtgatacgt acacttttcc
     2281 cggcaataac atacggcgtg acatcggctt caaatggcgt atagccgccc tgatgctcca
     2341 tcacttcctg attattgacc cacactttgc cgtaatgagt gaccgcatcg aaacgcagca
     2401 cgatacgctg gcctgcccaa cctttcggta taaagacttc gcgctgatac cagacgttgc
     2461 ccgcataatt acgaatatct gcatcggcga actgatcgtt aaaactgcct ggcacagcaa
     2521 ttgcccggct ttcttgtaac gcgctttccc accaacgctg atcaattcca cagttttcgc
     2581 gatccagact gaatgcccac aggccgtcga gttttttgat ttcacgggtt ggggtttcta
     2641 caggacggac catggtcgac tcctccttag ctggatctta gctcctgaaa atctcgacaa
     2701 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     2761 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     2821 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtaggc gcgccgaagt
     2881 tcctatactt tctagagaat aggaacttcg gaataggaac taaggaggat attcatatgg
     2941 accatggcta attcccatgt cagccgttaa gtgttcctgt gtcactgaaa attgctttga
     3001 gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac
     3061 cttaaaagct ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga
     3121 ggctatttaa gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt
     3181 gaaacatgag agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta
     3241 gttcgttaaa catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag
     3301 agcttagtac gtactatcaa caggttgaac tgcggatctt gcggccgcaa aaattaaaaa
     3361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3541 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3601 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3661 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3721 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3781 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3841 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3901 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3961 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4021 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4081 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4141 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4201 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4261 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4321 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4381 tttccccgaa aagtgccacc tgcatcgatg gccccccgat ggtagtgtgg ggtctcccca
     4441 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     4501 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     4561 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     4621 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtggc
     4681 cagtgccaag cttgcatgc
//
