LOCUS       sequence-252578-        7641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination low-copy number plasmid for expression of any 
            effector from avrPto promoter with C-terminal Cya tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7641)
  AUTHORS   Oh HS, Kvitko BH, Morello JE, Collmer A
  TITLE     Pseudomonas syringae lytic transglycosylases coregulated with the 
            type III secretion system contribute to the translocation of 
            effector proteins into plant cells.
  JOURNAL   J Bacteriol. 2007 Nov;189(22):8277-89. Epub 2007 Sep 7.
  PUBMED    17827286
REFERENCE   2  (bases 1 to 7641)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J 
            Bacteriol."; date: "2007-11"; volume: "189(22)"; pages: "8277-89. 
            Epub 2007 Sep 7"
COMMENT     Sequence Label: pCPP5371
FEATURES             Location/Qualifiers
     source          1..7641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(483..488,489..505,506..511))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        join(1024..1029,1030..1047,1048..1054)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1108..1767
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1174..1193)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2109..2414
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2293..2312
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2455..2579)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    3817..3924
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3833..3849
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3834..3850
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3884..3900)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3884..3900)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3932..3951)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3933..3951)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3961..3978)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3961..3977)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3970..3992)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4686..5348)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      5349..6120
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             complement(join(7402..7641,1..294))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag
       61 cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg
      121 ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga
      181 tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg agccacctaa
      241 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc
      301 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
      361 gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac
      421 caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt
      481 acagtttacg aaccgaacag gcttatgtca actgggttcg tgccttcatc cgtttccacg
      541 gtgtgcgtcc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
      601 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
      661 acagttttta tgcattaccg agtaacgcag ctgcgtttac ggcggacgca gccaaggcag
      721 cgttactgtg cgccaaactc aagtcagtga cgctttgatg tctaaacccg gcgccagcaa
      781 aattttgggg aattatctta taagcttgga accgatccgc tccctatgac cactcaagtg
      841 gcgtcataaa tgctgtacta aagagggtat atgcatgcgt ttgtacaaaa aagctgaacg
      901 agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata aaaaacagac
      961 tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcatt aggcacccca
     1021 ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg
     1081 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     1141 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1201 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1261 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1321 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1381 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1441 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1501 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1561 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1621 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1681 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1741 tgcgatgagt ggcagggcgg ggcgtaaaga tctggatccg gcttactaaa agccagataa
     1801 cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc
     1861 cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg
     1921 acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac
     1981 catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg
     2041 gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg
     2101 ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
     2161 tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca
     2221 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
     2281 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg
     2341 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
     2401 tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt cgaccatagt
     2461 gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt
     2521 aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtt
     2581 gatgggcagc aatcgcatca ggctggttac gcaaacgccg ccgaccggga gtctggcatc
     2641 cccgcagccg tactcgatgg catcaaggcc gtggcgaagg aaaaaaacgc cacattgatg
     2701 ttccgcctgg tcaaccccca ttccaccagc ctgattgccg aaggggtggc caccaaagga
     2761 ttgggcgtgc acgccaagtc gtccgattgg gggttgcagg cgggctacat tcccgtcaac
     2821 ccgaatcttt ccaaactgtt cggccgtgcg cccgaggtga tcgcgcgggc cgacaacgac
     2881 gtcaacagca gcctggcgca tggccatacc gcggtcgacc tgacgctgtc gaaagagcgg
     2941 cttgactatc tgcggcaagc gggcctggtc accggcatgg ccgatggcgt ggtcgcgagc
     3001 aaccacgcag gctacgagca gttcgagttt cgcgtgaagg aaacctcgga cgggcgctat
     3061 gccgtgcagt atcgccacaa gggcggcgac gatttcgagg cggtcaaggt gatcggcaat
     3121 gccgccggta ttccactgac ggcggatatc gacatgttcg ccattatgcc acatctgtcc
     3181 aacttccgcg actcggcgcg cagttcggtg accagcggcg attcggtgac cgattacctg
     3241 gcgcgcatgc ggcgggccgc cagcgaggcc acgggcggcc tggatcgcga acgcatcgac
     3301 ttgttgtgga aaatcgctcg cgccggcgcc cgttccgcag tgggcaccga ggcgcgtcgc
     3361 cagttccgct acgacggcga catgaatatc ggcgtgatca ccgatttcga gctggaagtg
     3421 cgcaatgcgc tgaacaggcg ggcgcacgcc gtcggcgcgc aggacgtggt ccagcatggc
     3481 actgagcaga acaatccttt cccggaggca gatgagaaga ttttcgtcgt atcggccacc
     3541 ggtgaaagcc agatgctcac gcgcgggcaa ctgaaggaat acattggcca gcagcgcagc
     3601 gagggctatg tcttctacga gaaccgtgca tacggcgtgg cggggaaaag cctgttcgac
     3661 gatgggctgg gagccgtgcc cggcgtgccg agcggacgtt cgaagttctc gccggatgta
     3721 ctggaaacgg tgccggcgtc acccggattg cggcggccgt cgctgggcgc agtggaacgc
     3781 caggattccg gctatgacag ctaagaattc gatatcggta ccgggccccc cctcgaggtc
     3841 gacggtatcg ataagcttga tatcgaattc ctgcagcccg ggggatccac tagttctaga
     3901 gcggccgcca ccgcggtgga gctccaattc gccctatagt gagtcgtatt acgcgcgctc
     3961 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4021 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4081 cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg
     4141 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgtactg
     4201 cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc
     4261 tggttgctac gcctgaataa gtgataataa gcggatgaat ggcagaaatt cgaaagcaaa
     4321 ttcgacccgg tcgtcggttc agggcagggt cgttaaatag ccgcttatgt ctattgctgg
     4381 tttaccggtt tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc
     4441 cagtttgctc aggctctccc cgtggaggta ataattgacg atatgatcat ttattctgcc
     4501 tcccagagcc tgataaaaac ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     4561 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     4621 cggcgaggcg gctacagccg atagtctgga acagcgcact tacgggttgc tgcgcaaccc
     4681 aagtgctacc ggcgcggcag cgtgacccgt gtcggcggct ccaacggctc gccatcgtcc
     4741 agaaaacacg gctcatcggg catcggcagg cgctgctgcc cgcgccgttc ccattcctcc
     4801 gtttcggtca aggctggcag gtctggttcc atgcccggaa tgccgggctg gctgggcggc
     4861 tcctcgccgg ggccggtcgg tagttgctgc tcgcccggat acagggtcgg gatgcggcgc
     4921 aggtcgccat gccccaacag cgattcgtcc tggtcgtcgt gatcaaccac cacggcggca
     4981 ctgaacaccg acaggcgcaa ctggtcgcgg ggctggcccc acgccacgcg gtcattgacc
     5041 acgtaggccg acacggtgcc ggggccgttg agcttcacga cggagatcca gcgctcggcc
     5101 accaagtcct tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag cttggaaagt
     5161 gtcttctggc tgaccaccac ggcgttctgg tggcccatct gcgccacgag gtgatgcagc
     5221 agcattgccg ccgtgggttt cctcgcaata agcccggccc acgcctcatg cgctttgcgt
     5281 tccgtttgca cccagtgacc gggcttgttc ttggcttgaa tgccgatttc tctggactgc
     5341 gtggccatgc ttatctccat gcggtagggg tgccgcacgg ttgcggcacc atgcgcaatc
     5401 agctgcaact tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg gcagtcaatt
     5461 acagattttc tttaacctac gcaatgagct attgcggggg gtgccgcaat gagctgttgc
     5521 gtacccccct tttttaagtt gttgattttt aagtctttcg catttcgccc tatatctagt
     5581 tctttggtgc ccaaagaagg gcacccctgc ggggttcccc cacgccttcg gcgcggctcc
     5641 ccctccggca aaaagtggcc cctccggggc ttgttgatcg actgcgcggc cttcggcctt
     5701 gcccaaggtg gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg ctcggggggc
     5761 aggcgggcgg gcttcgccct tcgactgccc ccactcgcat aggcttgggt cgttccaggc
     5821 gcgtcaaggc caagccgctg cgcggtcgct gcgcgagcct tgacccgcct tccacttggt
     5881 gtccaaccgg caagcgaagc gcgcaggccg caggccggag gcttttcccc agagaaaatt
     5941 aaaaaaattg atggggcaag gccgcaggcc gcgcagttgg agccggtggg tatgtggtcg
     6001 aaggctgggt agccggtggg caatccctgt ggtcaagctc gtgggcaggc gcagcctgtc
     6061 catcagcttg tccagcaggg ttgtccacgg gccgagcgaa gcgagccagc cggtggccgc
     6121 tcgcggccat cgtccacata tccacgggct ggcaagggag cgcagcgacc gcgcagggcg
     6181 aagcccggag agcaagcccg tagggcgccg cagccgccgt aggcggtcac gactttgcga
     6241 agcaaagtct agtgagtata ctcaagcatt gagtggcccg ccggaggcac cgccttgcgc
     6301 tgcccccgtc gagccggttg gacaccaaaa gggaggggca ggcatggcgg catacgcgat
     6361 catgcgatgc aagaagctgg cgaaaatggg caacgtggcg gccagtctca agcacgccta
     6421 ccgcgagcgc gagacgccca acgctgacgc cagcaggacg ccagagaacg agcactgggc
     6481 ggccagcagc accgatgaag cgatgggccg actgcgcgag ttgctgccag agaagcggcg
     6541 caaggacgct gtgttggcgg tcgagtacgt catgacggcc agcccggaat ggtggaagtc
     6601 ggccagccaa gaacagcagg cggcgttctt cgagaaggcg cacaagtggc tggcggacaa
     6661 gtacggggcg gatcgcatcg tgacggccag catccaccgt gacgaaacca gcccgcacat
     6721 gaccgcgttc gtggtgccgc tgacgcagga cggcaggctg tcggccaagg agttcatcgg
     6781 caacaaagcg cagatgaccc gcgaccagac cacgtttgcg gccgctgtgg ccgatctagg
     6841 gctgcaacgg ggcatcgagg gcagcaaggc acgtcacacg cgcattcagg cgttctacga
     6901 ggccctggag cggccaccag tgggccacgt caccatcagc ccgcaagcgg tcgagccacg
     6961 cgcctatgca ccgcagggat tggccgaaaa gctgggaatc tcaaagcgcg ttgagacgcc
     7021 ggaagccgtg gccgaccggc tgacaaaagc ggttcggcag gggtatgagc ctgccctaca
     7081 ggccgccgca ggagcgcgtg agatgcgcaa gaaggccgat caagcccaag agacggcccg
     7141 agaccttcgg gagcgcctga agcccgttct ggacgccctg gggccgttga atcgggatat
     7201 gcaggccaag gccgccgcga tcatcaaggc cgtgggcgaa aagctgctga cggaacagcg
     7261 ggaagtccag cgccagaaac aggcccagcg ccagcaggaa cgcgggcgcg cacatttccc
     7321 cgaaaagtgc cacctgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     7381 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     7441 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     7501 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     7561 cgcggtggca atgccctgcc tccggtgctc tccggagact gcgagatcat agatatagat
     7621 ctcactacgc ggctgctcaa a
//
