LOCUS       40924_6087        7187 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBBR5pemIK-GW, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7187)
  AUTHORS   Burbank L, Stenger DC.
  TITLE     Plasmid vectors for Xylella fastidiosa utilizing a toxin-antitoxin 
            system for plasmid stability in the absence of antibiotic selection
  JOURNAL   Phytopathology (2016) In press
  PUBMED    27088393
REFERENCE   2  (bases 1 to 7187)
  AUTHORS   Burbank LP, Stenger DC.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2016) Crop Diseases Pests and Genetics Research 
            Unit, USDA Agricultural Research Service, 9611 S Riverbend Ave, 
            Parlier, CA 93648, USA
REFERENCE   3  (bases 1 to 7187)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7187)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Phytopathology (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-APR-2016) Crop Diseases Pests and Genetics Research Unit, USDA 
            Agricultural Research Service, 9611 S Riverbend Ave, Parlier, CA 
            93648, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             538..1197
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     1909..1925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1935..1953
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1986..2002
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2010..2134
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2159..2189
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2243..2899
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3244..3546
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3590..3714)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3805..4068
                     /codon_start=1
                     /gene="pemI"
                     /product="antitoxin"
                     /label=pemI
                     /protein_id="ANA75337.1"
                     /translation="MHTTNLRKVGGSIMLAVPPAFLDQLHLEVGATVGLAVTDGRLVIE
                     PVLHPQYTLDQLLAEAEASGAYPLPPEEREWVDAPAVGRELI"
     gene            3805..4068
                     /gene="pemI"
                     /label=pemI
     CDS             4065..4391
                     /codon_start=1
                     /gene="pemK"
                     /product="mRNA interferase"
                     /label=pemK
                     /protein_id="ANA75336.1"
                     /translation="MKRGDVYMVDLEPTAGHEQRGHRPVVVISSERFNRLTSCPVILPI
                     TNGGEFASRLGFAVELLGTITTGVVRCDQPRALDLLARNARKVESLPPNILADVLAKAV
                     TIFQ"
     gene            4065..4391
                     /gene="pemK"
                     /label=pemK
     primer_bind     complement(4452..4468)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4498..4516)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4537..4553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4561..4577)
                     /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(4585..4615)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4630..4651)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4921..4949
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5138..5668
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      6953..7187
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 tggacaccaa gtggaaggcg ggtcaaggct cgcgcagcga ccgcgcagcg gcttggcctt
       61 gacgcgcctg gaacgaccca agcctatgcg agtgggggca gtcgaaaggc gaagcccgcc
      121 cgcctgcccc ccgagcctca cggcggcgag tgcgggggtt ccaagggggc agcgccacct
      181 tgggcaaggc cgaaggccgc gcagtcgatc aacaagcccc ggaggggcca ctttttgccg
      241 gagggggagc cgcgccgaag gcgtggggga accccgcagg ggtgcccttc tttgggcacc
      301 aaagaactag atatagggcg aaatgcgaaa gacttaaaaa tcaacaactt aaaaaagggg
      361 ggtacgcaac agctcattgc ggcacccccc gcaatagctc attgcgtagg ttaaagaaaa
      421 tctgtaattg actgccactt ttacgcaacg cataattgtt gtcgcgctgc cgaaaagttg
      481 cagctgattg cgcatggtgc cgcaaccgtg cggcacccct accgcatgga gataagcatg
      541 gccacgcagt ccagagaaat cggcattcaa gccaagaaca agcccggtca ctgggtgcaa
      601 acggaacgca aagcgcatga ggcgtgggcc gggcttattg cgaggaaacc cacggcggca
      661 atgctgctgc atcacctcgt ggcgcagatg ggccaccaga acgccgtggt ggtcagccag
      721 aagacacttt ccaagctcat cggacgttct ttgcggacgg tccaatacgc agtcaaggac
      781 ttggtggccg agcgctggat ctccgtcgtg aagctcaacg gccccggcac cgtgtcggcc
      841 tacgtggtca atgaccgcgt ggcgtggggc cagccccgcg accagttgcg cctgtcggtg
      901 ttcagtgccg ccgtggtggt tgatcacgac gaccaggacg aatcgctgtt ggggcatggc
      961 gacctgcgcc gcatcccgac cctgtatccg ggcgagcagc aactaccgac cggccccggc
     1021 gaggagccgc ccagccagcc cggcattccg ggcatggaac cagacctgcc agccttgacc
     1081 gaaacggagg aatgggaacg gcgcgggcag cagcgcctgc cgatgcccga tgagccgtgt
     1141 tttctggacg atggcgagcc gttggagccg ccgacacggg tcacgctgcc gcgccggtag
     1201 cacttgggtt gcgcagcaac ccgtaagtgc gctgttccag actatcggct gtagccgcct
     1261 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     1321 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccgtttt tatcaggctc
     1381 tgggaggcag aataaatgat catatcgtca attattacct ccacggggag agcctgagca
     1441 aactggcctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     1501 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     1561 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     1621 aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
     1681 catcgcagta cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1741 tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg
     1801 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     1861 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     1921 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctccac cgcggtggcg
     1981 gccgctctag aactagtgga tcccccatca caagtttgta caaaaaagct gaacgagaaa
     2041 cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa cagactacat
     2101 aatactgtaa aacacaacat atccagtcac tatggcggcc gcattaggca ccccaggctt
     2161 tacactttat gcttccggct cgtataatgt gtggattttg agttaggatc cgtcgagatt
     2221 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     2281 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     2341 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     2401 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     2461 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     2521 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     2581 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     2641 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     2701 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     2761 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     2821 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     2881 tgagtggcag ggcggggcgt aaacgcgtgg atccggctta ctaaaagcca gataacagta
     2941 tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag
     3001 tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc
     3061 tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc
     3121 agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg
     3181 ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt
     3241 gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
     3301 gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca
     3361 cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa
     3421 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa
     3481 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
     3541 ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc atagtgactg
     3601 gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat
     3661 attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag tggtgatggg
     3721 ctgcaggaat tcgattttcg ctataagcgt ataaatggag gaattatatt atactttaat
     3781 ataacttaac gaaagaagcc catcatgcac accacaaacc ttcgtaaagt cggtggatcg
     3841 atcatgctgg ccgttccccc cgctttcctt gatcagcttc atcttgaggt cggcgcaacg
     3901 gtcggcttag ccgtcacaga cggtcgtctg gtcattgagc cagtcctaca tcctcagtac
     3961 acgctggatc aactgctggc cgaagcggaa gcctccgggg cttacccgtt gccgcccgag
     4021 gaacgtgaat gggtcgatgc tccagcagta ggacgcgaac tgatatgaaa cgcggcgatg
     4081 tttacatggt cgatctggag ccaaccgccg gacacgaaca acgcggtcac cgtccggtgg
     4141 ttgttatctc cagtgagcga ttcaatcggc ttacctcttg tccggtgatc cttccaatca
     4201 caaacggcgg cgagttcgcc tcccgcctcg ggttcgctgt ggaattacta ggtacgatta
     4261 caacgggcgt cgttcgatgt gatcaaccac gtgcgctcga cttactcgct cgcaatgccc
     4321 gcaaagtgga aagcctacca ccaaacatct tggccgatgt cttggcaaaa gctgtcacca
     4381 tcttccagta atttttgggg caagttcagg ccactgcaac caatcactag tgaattcgat
     4441 atcaagctta tcgataccgt cgacctcgag ggggggcccg gtacccagct tttgttccct
     4501 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4561 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4621 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4681 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4741 tttgcgtatt gggcgcatgc ataaaaactg ttgtaattca ttaagcattc tgccgacatg
     4801 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     4861 ttgcgtataa tatttgccca tggacgcaca ccgtggaaac ggatgaaggc acgaacccag
     4921 ttgacataag cctgttcggt tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac
     4981 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     5041 tgttatgact gtttttttgt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     5101 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gcaacgatgt
     5161 tacgcagcag ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg
     5221 cacatgtagg ctcggccctg accaagtcaa atccatgcgg gctgctcttg atcttttcgg
     5281 tcgtgagttc ggagacgtag ccacctactc ccaacatcag ccggactccg attacctcgg
     5341 gaacttgctc cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt
     5401 tggcgctctc gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg agatctatat
     5461 ctatgatctc gcagtctccg gagagcaccg gaggcagggc attgccaccg cgctcatcaa
     5521 tctcctcaag catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta
     5581 cggtgacgat cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca
     5641 ctttgatatc gacccaagta ccgccaccta acaattcgtt caagccgaga tcggcttccc
     5701 ggccgcggag ttgttcggta aattgtcaca acgccgcagg tggcactttt cggggaaatg
     5761 tgcgcgcccg cgttcctgct ggcgctgggc ctgtttctgg cgctggactt cccgctgttc
     5821 cgtcagcagc ttttcgccca cggccttgat gatcgcggcg gccttggcct gcatatcccg
     5881 attcaacggc cccagggcgt ccagaacggg cttcaggcgc tcccgaaggt ctcgggccgt
     5941 ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg cctgtagggc
     6001 aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt ccggcgtctc
     6061 aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg cgcgtggctc
     6121 gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg cctcgtagaa
     6181 cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca gccctagatc
     6241 ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt tgccgatgaa
     6301 ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg tcatgtgcgg
     6361 gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt acttgtccgc
     6421 cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg ccgacttcca
     6481 ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct tgcgccgctt
     6541 ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg ccgcccagtg
     6601 ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc ggtaggcgtg
     6661 cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca tgatcgcgta
     6721 tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg cagcgcaagg
     6781 cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg cttcgcaaag
     6841 tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc ttcgccctgc
     6901 gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc gagcggccac
     6961 cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga tggacaggct
     7021 gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc ttcgaccaca
     7081 tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt ttaattttct
     7141 ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggt
//
