pBBR5pemIK-GW vector (V009252)

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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"