LOCUS       sequence-252502-        6464 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 HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6464)
  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 6464)
  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: pCPP5372
FEATURES             Location/Qualifiers
     source          1..6464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..125
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(150..155,156..173,174..180)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             234..893
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYLPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(300..319)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1235..1540
                     /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     1419..1438
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1581..1705)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1713..1739
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1762..1869
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1778..1794
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1779..1795
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1829..1845)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1829..1845)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1878..1896)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1906..1923)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1906..1922)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1915..1937)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2631..3293)
                     /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      3294..4065
                     /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(5347..5880)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6069..6074,6075..6091,6092..6097))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta
       61 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
      121 ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
      181 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
      241 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
      301 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
      361 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
      421 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
      481 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
      541 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
      601 atgtggcgtg ttacggtgaa aacctggcct atctccctaa agggtttatt gagaatatgt
      661 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
      721 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
      781 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
      841 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaagatctg
      901 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
      961 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     1021 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     1081 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     1141 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     1201 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     1261 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     1321 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     1381 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     1441 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     1501 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     1561 cagccagtct gcaggtcgac catagtgact ggatatgttg tgtcttacag tattatgtag
     1621 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     1681 gttcagcttt cttgtacaaa gtggttgatg tgtacccata cgacgtccca gactacgctt
     1741 aagggactcc ctatagtgag tggtaccggg ccccccctcg aggtcgacgg tatcgataag
     1801 cttgatatcg aattcctgca gcccggggga tccactagtt ctagagcggc cgccaccgcg
     1861 gtggagctcc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     1921 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1981 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2041 gcgcagcctg aatggcgaat ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     2101 aatttttgtt aaatcagctc attttttaac caataggccg tactgcgatg agtggcaggg
     2161 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     2221 aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga cccggtcgtc
     2281 ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac cggtttattg
     2341 actaccggaa gcagtgtgac cgtgtgcttc tcaaatgcct gaggccagtt tgctcaggct
     2401 ctccccgtgg aggtaataat tgacgatatg atcatttatt ctgcctccca gagcctgata
     2461 aaaacggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag gtggcccggc
     2521 tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg aggcggctac
     2581 agccgatagt ctggaacagc gcacttacgg gttgctgcgc aacccaagtg ctaccggcgc
     2641 ggcagcgtga cccgtgtcgg cggctccaac ggctcgccat cgtccagaaa acacggctca
     2701 tcgggcatcg gcaggcgctg ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct
     2761 ggcaggtctg gttccatgcc cggaatgccg ggctggctgg gcggctcctc gccggggccg
     2821 gtcggtagtt gctgctcgcc cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc
     2881 aacagcgatt cgtcctggtc gtcgtgatca accaccacgg cggcactgaa caccgacagg
     2941 cgcaactggt cgcggggctg gccccacgcc acgcggtcat tgaccacgta ggccgacacg
     3001 gtgccggggc cgttgagctt cacgacggag atccagcgct cggccaccaa gtccttgact
     3061 gcgtattgga ccgtccgcaa agaacgtccg atgagcttgg aaagtgtctt ctggctgacc
     3121 accacggcgt tctggtggcc catctgcgcc acgaggtgat gcagcagcat tgccgccgtg
     3181 ggtttcctcg caataagccc ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag
     3241 tgaccgggct tgttcttggc ttgaatgccg atttctctgg actgcgtggc catgcttatc
     3301 tccatgcggt aggggtgccg cacggttgcg gcaccatgcg caatcagctg caacttttcg
     3361 gcagcgcgac aacaattatg cgttgcgtaa aagtggcagt caattacaga ttttctttaa
     3421 cctacgcaat gagctattgc ggggggtgcc gcaatgagct gttgcgtacc cccctttttt
     3481 aagttgttga tttttaagtc tttcgcattt cgccctatat ctagttcttt ggtgcccaaa
     3541 gaagggcacc cctgcggggt tcccccacgc cttcggcgcg gctccccctc cggcaaaaag
     3601 tggcccctcc ggggcttgtt gatcgactgc gcggccttcg gccttgccca aggtggcgct
     3661 gcccccttgg aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc
     3721 gcccttcgac tgcccccact cgcataggct tgggtcgttc caggcgcgtc aaggccaagc
     3781 cgctgcgcgg tcgctgcgcg agccttgacc cgccttccac ttggtgtcca accggcaagc
     3841 gaagcgcgca ggccgcaggc cggaggcttt tccccagaga aaattaaaaa aattgatggg
     3901 gcaaggccgc aggccgcgca gttggagccg gtgggtatgt ggtcgaaggc tgggtagccg
     3961 gtgggcaatc cctgtggtca agctcgtggg caggcgcagc ctgtccatca gcttgtccag
     4021 cagggttgtc cacgggccga gcgaagcgag ccagccggtg gccgctcgcg gccatcgtcc
     4081 acatatccac gggctggcaa gggagcgcag cgaccgcgca gggcgaagcc cggagagcaa
     4141 gcccgtaggg cgccgcagcc gccgtaggcg gtcacgactt tgcgaagcaa agtctagtga
     4201 gtatactcaa gcattgagtg gcccgccgga ggcaccgcct tgcgctgccc ccgtcgagcc
     4261 ggttggacac caaaagggag gggcaggcat ggcggcatac gcgatcatgc gatgcaagaa
     4321 gctggcgaaa atgggcaacg tggcggccag tctcaagcac gcctaccgcg agcgcgagac
     4381 gcccaacgct gacgccagca ggacgccaga gaacgagcac tgggcggcca gcagcaccga
     4441 tgaagcgatg ggccgactgc gcgagttgct gccagagaag cggcgcaagg acgctgtgtt
     4501 ggcggtcgag tacgtcatga cggccagccc ggaatggtgg aagtcggcca gccaagaaca
     4561 gcaggcggcg ttcttcgaga aggcgcacaa gtggctggcg gacaagtacg gggcggatcg
     4621 catcgtgacg gccagcatcc accgtgacga aaccagcccg cacatgaccg cgttcgtggt
     4681 gccgctgacg caggacggca ggctgtcggc caaggagttc atcggcaaca aagcgcagat
     4741 gacccgcgac cagaccacgt ttgcggccgc tgtggccgat ctagggctgc aacggggcat
     4801 cgagggcagc aaggcacgtc acacgcgcat tcaggcgttc tacgaggccc tggagcggcc
     4861 accagtgggc cacgtcacca tcagcccgca agcggtcgag ccacgcgcct atgcaccgca
     4921 gggattggcc gaaaagctgg gaatctcaaa gcgcgttgag acgccggaag ccgtggccga
     4981 ccggctgaca aaagcggttc ggcaggggta tgagcctgcc ctacaggccg ccgcaggagc
     5041 gcgtgagatg cgcaagaagg ccgatcaagc ccaagagacg gcccgagacc ttcgggagcg
     5101 cctgaagccc gttctggacg ccctggggcc gttgaatcgg gatatgcagg ccaaggccgc
     5161 cgcgatcatc aaggccgtgg gcgaaaagct gctgacggaa cagcgggaag tccagcgcca
     5221 gaaacaggcc cagcgccagc aggaacgcgg gcgcgcacat ttccccgaaa agtgccacct
     5281 gcggcgttgt gacaatttac cgaacaactc cgcggccggg aagccgatct cggcttgaac
     5341 gaattgttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc
     5401 caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac
     5461 ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc
     5521 ctgcctccgg tgctctccgg agactgcgag atcatagata tagatctcac tacgcggctg
     5581 ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc
     5641 agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg
     5701 atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg
     5761 catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc
     5821 acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat
     5881 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
     5941 cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc
     6001 cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact
     6061 tgcattacag tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt
     6121 tccacggtgt gcgtccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     6181 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     6241 attacaacag tttttatgca ttaccgagta acgcagctgc gtttacggcg gacgcagcca
     6301 aggcagcgtt actgtgcgcc aaactcaagt cagtgacgct ttgatgtcta aacccggcgc
     6361 cagcaaaatt ttggggaatt atcttataag cttggaaccg atccgctccc tatgaccact
     6421 caagtggcgt cataaatgct gtactaaaga gggtatatgc atgc
//
