LOCUS       Exported                7187 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBBR5pemIK-GW, complete sequence.
ACCESSION   KX036764
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7187
                     /organism="Cloning vector pBBR5pemIK-GW"
                     /mol_type="other DNA"
                     /db_xref="taxon:1840503"
     rep_origin      526..1199
     CDS             538..1200
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /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"
     misc_recomb     2010..2134
                     /label=attR
                     /note="attR"
     protein_bind    2010..2134
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /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..2902
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="chloramphenicol resistance"
                     /protein_id="ANA75339.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            2243..2902
                     /gene="cat"
                     /label=cat
     CDS             3244..3549
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB killer protein"
                     /label=ccdB
                     /protein_id="ANA75338.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            3244..3549
                     /gene="ccdB"
                     /label=ccdB
     protein_bind    complement(3590..3714)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     3613..3690
                     /label=attR
                     /note="attR"
     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    4561..4577
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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    4630..4651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4921..4949
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5138..5671
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin-3-acetyltransferase"
                     /label=aacC1
                     /note="gentamicin resistance"
                     /protein_id="ANA75335.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            5138..5671
                     /gene="aacC1"
                     /label=aacC1
     CDS             complement(5680..7122)
                     /codon_start=1
                     /gene="mob"
                     /product="mobilization protein"
                     /label=mob
                     /protein_id="ANA75334.1"
                     /translation="MGQGRRPRSWSRWVCGRRLGSRWAIPVVKLVGRRSLSISLSSRVV
                     HGPSEASQPVAARGHRPHIHGLARERSDRAGRSPESKPVGRRSRRRRSRLCEAKSSEYT
                     QALSGPPEAPPCAAPVEPVGHQKGGAGMAAYAIMRCKKLAKMGNVAASLKHAYRERETP
                     NADASRTPENEHWAASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQ
                     QAAFFEKAHKWLADKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQ
                     MTRDQTTFAAAVADLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYA
                     PQGLAEKLGISKRVETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDL
                     RERLKPVLDALGPLNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPE
                     KCHLRRCDNLPNNSAAGKPISA"
     gene            complement(5680..7122)
                     /gene="mob"
                     /label=mob
     rep_origin      6953..537
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"