LOCUS       40924_13160        7567 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCPP5333, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7567)
  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 7567)
  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 7567)
  AUTHORS   Kvitko BH, Ramos AR, Morello JE, Oh H-S., Collmer A.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUL-2007) Plant Pathology, Cornell University, 334 
            Plant Science, Ithaca, NY 14853, USA
REFERENCE   4  (bases 1 to 7567)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7567)
  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 
            (09-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..7567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..45)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(53..69)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(77..107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(122..143)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(431..1019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1275..1805)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1994..2022)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(2040..2144)
                     /label=AmpR promoter
     misc_feature    2234..2251
                     /label=SCE
                     /note="SCE"
     CDS             complement(2476..3894)
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        complement(3895..4340)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     oriT            complement(4714..4823)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(5244..5271)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(5363..5449)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     5799..5815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5860..5979
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6009..6039
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             6093..6749
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             7094..7396
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(7440..7564)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gggtaccgag ctcgaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc
       61 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      121 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      181 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      241 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      301 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      361 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      421 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      481 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      541 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      601 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      661 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      721 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      781 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      841 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
      901 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      961 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1021 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1081 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1141 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacaat cgatgcgaat
     1201 tggccgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc gatctcggct
     1261 tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt
     1321 atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag
     1381 atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag cgcggtggca
     1441 atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat ctcactacgc
     1501 ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg
     1561 aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta atcggagtcc
     1621 ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag atcaagagca
     1681 gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat gcccatactt
     1741 gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctgcgt
     1801 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
     1861 ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac
     1921 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
     1981 ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aattcggttg aatactcata
     2041 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     2101 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     2161 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     2221 atcacgaggc ccttagggat aacagggtaa taggcccttt cgtctcgcgc gtttcggtga
     2281 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2341 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2401 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata atcggcattt
     2461 tcttttgcgt ttttatttgt taactgttaa ttgtccttgt tcaaggatgc tgtctttgac
     2521 aacagatgtt ttcttgcctt tgatgttcag caggaagcta ggcgcaaacg ttgattgttt
     2581 gtctgcgtag aatcctctgt ttgtcatata gcttgtaatc acgacattgt ttcctttcgc
     2641 ttgaggtaca gcgaagtgtg agtaagtaaa ggttacatcg ttaggatcaa gatccatttt
     2701 taacacaagg ccagttttgt tcagcggctt gtatgggcca gttaaagaat tagaaacata
     2761 accaagcatg taaatatcgt tagacgtaat gccgtcaatc gtcatttttg atccgcggga
     2821 gtcagtgaac agataccatt tgccgttcat tttaaagacg ttcgcgcgtt caatttcatc
     2881 tgttactgtg ttagatgcaa tcagcggttt catcactttt ttcagtgtgt aatcatcgtt
     2941 tagctcaatc ataccgagag cgccgtttgc taactcagcc gtgcgttttt tatcgctttg
     3001 cagaagtttt tgactttctt gacggaagaa tgatgtgctt ttgccatagt atgctttgtt
     3061 aaataaagat tcttcgcctt ggtagccatc ttcagttcca gtgtttgctt caaatactaa
     3121 gtatttgtgg cctttatctt ctacgtagtg aggatctctc agcgtatggt tgtcgcctga
     3181 gctgtagttg ccttcatcga tgaactgctg tacattttga tacgtttttc cgtcaccgtc
     3241 aaagattgat ttataatcct ctacaccgtt gatgttcaaa gagctgtctg atgctgatac
     3301 gttaacttgt gcagttgtca gtgtttgttt gccgtaatgt ttaccggaga aatcagtgta
     3361 gaataaacgg atttttccgt cagatgtaaa tgtggctgaa cctgaccatt cttgtgtttg
     3421 gtcttttagg atagaatcat ttgcatcgaa tttgtcgctg tctttaaaga cgcggccagc
     3481 gtttttccag ctgtcaatag aagtttcgcc gactttttga tagaacatgt aaatcgatgt
     3541 gtcatccgca tttttaggat ctccggctaa tgcaaagacg atgtggtagc cgtgatagtt
     3601 tgcgacagtg ccgtcagcgt tttgtaatgg ccagctgtcc caaacgtcca ggccttttgc
     3661 agaagagata tttttaattg tggacgaatc gaactcagga acttgatatt tttcattttt
     3721 ttgctgttca gggatttgca gcatatcatg gcgtgtaata tgggaaatgc cgtatgtttc
     3781 cttatatggc ttttggttcg tttctttcgc aaacgcttga gttgcgcctc ctgccagcag
     3841 tgcggtagta aaggttaata ctgttgcttg ttttgcaaac tttttgatgt tcatcgttca
     3901 tgtctccttt tttatgtact gtgttagcgg tctgcttctt ccagccctcc tgtttgaaga
     3961 tggcaagtta gttacgcaca ataaaaaaag acctaaaata tgtaaggggt gacgccaaag
     4021 tatacacttt gccctttaca cattttaggt cttgcctgct ttatcagtaa caaacccgcg
     4081 cgatttactt ttcgacctca ttctattaga ctctcgtttg gattgcaact ggtctatttt
     4141 cctcttttgt ttgatagaaa atcataaaag gatttgcaga ctacgggcct aaagaactaa
     4201 aaaatctatc tgtttctttt cattctctgt attttttata gtttctgttg catgggcata
     4261 aagttgcctt tttaatcaca attcagaaaa tatcataata tctcatttca ctaaataata
     4321 gtgaacggca ggtatatgtg atgggttaaa aaggatcgat cctctagcta gagtcgatct
     4381 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
     4441 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
     4501 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
     4561 aggccgacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
     4621 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     4681 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     4741 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     4801 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     4861 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     4921 ctataatgac cccgaagcag ggttatgcag cggaaaagcg ctgcttccct gctgttttgt
     4981 ggaatatcta ccgactggaa acaggcaaat gcaggaaatt actgaactga ggggacaggc
     5041 gagagacgat gccaaagagc tacaccgacg agctggccga gtgggttgaa tcccgcgcgg
     5101 ccaagaagcg ccggcgtgat gaggctgcgg ttgcgttcct ggcggtgagg gcggatgtcg
     5161 atatgcgtaa ggagaaaata ccgcatcagg cgcatatttg aatgtattta gaaaaataaa
     5221 caaaaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt
     5281 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa
     5341 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     5401 agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag
     5461 ttccctactc tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct
     5521 gagttcggca tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt
     5581 atcagaccgc ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg
     5641 ccaaaacagc caagctcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5701 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5761 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     5821 gcttgcatgc ctgcaggtcg actctagagg atccccatca caagtttgta caaaaaagct
     5881 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa
     5941 cagactacat aatactgtaa aacacaacat atccagtcat attggcggcc gcattaggca
     6001 ccccaggctt tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc
     6061 cgtcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     6121 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     6181 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     6241 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     6301 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     6361 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     6421 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     6481 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
     6541 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     6601 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     6661 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     6721 agtactgcga tgagtggcag ggcggggcgt aaacgcgtgg atccggctta ctaaaagcca
     6781 gataacagta tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt
     6841 atacccgaag tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga
     6901 cagcgacagc tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc
     6961 acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag
     7021 gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac
     7081 aggggctggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
     7141 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct
     7201 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat
     7261 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat
     7321 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
     7381 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc
     7441 atagtgactg gatatgttgt gttttacagc attatgtagt ctgtttttta tgcaaaatct
     7501 aatttaatat attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag
     7561 tggtgat
//
