LOCUS       62056_3730        7368 bp DNA     circular SYN 06-DEC-2019
DEFINITION  Cloning vector pBBR4pemIK-GW, complete sequence.
ACCESSION   MN044102
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  AUTHORS   Burbank L, Wei W.
  TITLE     Broad-Host-Range Plasmids for Constitutive and Inducible Gene 
            Expression in the Absence of Antibiotic Selection
  JOURNAL   Microbiol Resour Announc 8 (36), e00769-19 (2019)
  PUBMED    31488530
REFERENCE   2  (bases 1 to 7368)
  AUTHORS   Burbank LP, Wei W.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2019) Crop Diseases Pests and Genetics Research 
            Unit, USDA Agricultural Research Service, 9611 S Riverbend Ave, 
            Parlier, CA 93648, USA
REFERENCE   3  (bases 1 to 7368)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microbiol 
            Resour Announc"; date: "2019"; volume: "8"; issue: "36"; pages: 
            "e00769-19"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUN-2019) Crop Diseases Pests and Genetics Research Unit, USDA 
            Agricultural Research Service, 9611 S Riverbend Ave, Parlier, CA 
            93648, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..770
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             771..1430
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     primer_bind     2140..2156
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2166..2184
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2217..2233
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2293..2655)
                     /codon_start=1
                     /transl_table=11
                     /product="mRNA interferase PemK"
                     /label=mRNA interferase PemK
                     /protein_id="QDJ95328.1"
                     /translation="MGRCSSSRTRTDMKRGDVYMVDLEPTAGHEQRGHRPVVVISSERF
                     NRLTSCPVILPITNGGEFASRLGFAVELLGTITTGVVRCDQPRALDLLARNARKVESLP
                     PNILADVLAKAVTIFQ"
     CDS             complement(2616..2879)
                     /codon_start=1
                     /transl_table=11
                     /product="antitoxin PemI"
                     /label=antitoxin PemI
                     /protein_id="QDJ95329.1"
                     /translation="MHTTNLRKVGGSIMLAVPPAFLDQLHLEVGATVGLAVTDGRLVIE
                     PVLHPQYTLDQLLAEAEASGAYPLPPEEREWVDAPAVGRELI"
     protein_bind    2965..3089
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3114..3144
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3198..3854
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4199..4501
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(4545..4669)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4685..4701)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4731..4749)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4770..4786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4794..4810)
                     /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(4818..4848)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4863..4884)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5112..5183
                     /label=AmpR promoter
     CDS             5184..6041
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga caagctgatg
       61 gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct acccagcctt
      121 cgaccacata cccaccggct ccaactgcgc ggcctgcggc cttgccccat caattttttt
      181 aattttctct ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg ccggttggac
      241 accaagtgga aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg gccttgacgc
      301 gcctggaacg acccaagcct atgcgagtgg gggcagtcga aggcgaagcc cgcccgcctg
      361 ccccccgagc ctcacggcgg cgagtgcggg ggttccaagg gggcagcgcc accttgggca
      421 aggccgaagg ccgcgcagtc gatcaacaag ccccggaggg gccacttttt gccggagggg
      481 gagccgcgcc gaaggcgtgg gggaaccccg caggggtgcc cttctttggg caccaaagaa
      541 ctagatatag ggcgaaatgc gaaagactta aaaatcaaca acttaaaaaa ggggggtacg
      601 caacagctca ttgcggcacc ccccgcaata gctcattgcg taggttaaag aaaatctgta
      661 attgactgcc acttttacgc aacgcataat tgttgtcgcg ctgccgaaaa gttgcagctg
      721 attgcgcatg gtgccgcaac cgtgcggcac cctaccgcat ggagataagc atggccacgc
      781 agtccagaga aatcggcatt caagccaaga acaagcccgg tcactgggtg caaacggaac
      841 gcaaagcgca tgaggcgtgg gccgggctta ttgcgaggaa acccacggcg gcaatgctgc
      901 tgcatcacct cgtggcgcag atgggccacc agaacgccgt ggtggtcagc cagaagacac
      961 tttccaagct catcggacgt tctttgcgga cggtccaata cgcagtcaag gacttggtgg
     1021 ccgagcgctg gatctccgtc gtgaagctca acggccccgg caccgtgtcg gcctacgtgg
     1081 tcaatgaccg cgtggcgtgg ggccagcccc gcgaccagtt gcgcctgtcg gtgttcagtg
     1141 ccgccgtggt ggttgatcac gacgaccagg acgaatcgct gttggggcat ggcgacctgc
     1201 gccgcatccc gaccctgtat ccgggcgagc agcaactacc gaccggcccc ggcgaggagc
     1261 cgcccagcca gcccggcatt ccgggcatgg aaccagacct gccagccttg accgaaacgg
     1321 aggaatggga acggcgcggg cagcagcgcc tgccgatgcc cgatgagccg tgttttctgg
     1381 acgatggcga gccgttggag ccgccgacac gggtcacgct gccgcgccgg tagcacttgg
     1441 gttgcgcagc aacccgtaag tgcgctgttc cagactatcg gctgtagccg cctcgccgcc
     1501 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     1561 tgaatggaag ccggcggcac ctcgctaacg gattcaccgt ttttatcagg ctctgggagg
     1621 cagaataaat gatcatatcg tcaattatta cctccacggg gagagcctga gcaaactggc
     1681 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     1741 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt
     1801 tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt agcaccaggc
     1861 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     1921 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1981 atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2041 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2101 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     2161 gcgcgtaata cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta
     2221 gaactagtgg atcccccggg ctgcaggaat tcactagtga ttggttgcag tggcctgaac
     2281 ttgccccaaa aattactgga agatggtgac agcttttgcc aagacatcgg ccaagatgtt
     2341 tggtggtagg ctttccactt tgcgggcatt gcgagcgagt aagtcgagcg cacgtggttg
     2401 atcacatcga acgacgcccg ttgtaatcgt acctagtaat tccacagcga acccgaggcg
     2461 ggaggcgaac tcgccgccgt ttgtgattgg aaggatcacc ggacaagagg taagccgatt
     2521 gaatcgctca ctggagataa caaccaccgg acggtgaccg cgttgttcgt gtccggcggt
     2581 tggctccaga tcgaccatgt aaacatcgcc gcgtttcata tcagttcgcg tcctactgct
     2641 ggagcatcga cccattcacg ttcctcgggc ggcaacgggt aagccccgga ggcttccgct
     2701 tcggccagca gttgatccag cgtgtactga ggatgtagga ctggctcaat gaccagacga
     2761 ccgtctgtga cggctaagcc gaccgttgcg ccgacctcaa gatgaagctg atcaaggaaa
     2821 gcggggggaa cggccagcat gatcgatcca ccgactttac gaaggtttgt ggtgtgcatg
     2881 atgggcttct ttcgttaagt tatattaaag tataatataa ttcctccatt tatacgctta
     2941 tagcgaaaat cgaattcgat atcaacaagt ttgtacaaaa aagctgaacg agaaacgtaa
     3001 aatgatataa atatcaatat attaaattag attttgcata aaaaacagac tacataatac
     3061 tgtaaaacac aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac
     3121 tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag
     3181 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     3241 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     3301 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     3361 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta
     3421 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     3481 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     3541 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     3601 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     3661 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     3721 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     3781 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     3841 ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt
     3901 atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt
     3961 caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca
     4021 gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat
     4081 gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag
     4141 gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat
     4201 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     4261 gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct
     4321 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     4381 gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc
     4441 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     4501 ataaatgtca ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat
     4561 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
     4621 tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtt gatatcaagc
     4681 ttatcgatac cgtcgacctc gagggggggc ccggtaccca gcttttgttc cctttagtga
     4741 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4801 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4861 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4921 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4981 attgggcgca tgcataaaaa ctgttgtaat tcattaagca ttctgccgac atggaagcca
     5041 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     5101 taatatttgc ccattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5161 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5221 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5281 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5341 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5401 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5461 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5521 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5581 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5641 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5701 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5761 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5821 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5881 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5941 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6001 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6061 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6121 agatcctttt tgataatctc atgaccaaaa tccaggtggc acttttcggg gaaatgtgcg
     6181 cgcccgcgtt cctgctggcg ctgggcctgt ttctggcgct ggacttcccg ctgttccgtc
     6241 agcagctttt cgcccacggc cttgatgatc gcggcggcct tggcctgcat atcccgattc
     6301 aacggcccca gggcgtccag aacgggcttc aggcgctccc gaaggtctcg ggccgtctct
     6361 tgggcttgat cggccttctt gcgcatctca cgcgctcctg cggcggcctg tagggcaggc
     6421 tcatacccct gccgaaccgc ttttgtcagc cggtcggcca cggcttccgg cgtctcaacg
     6481 cgctttgaga ttcccagctt ttcggccaat ccctgcggtg cataggcgcg tggctcgacc
     6541 gcttgcgggc tgatggtgac gtggcccact ggtggccgct ccagggcctc gtagaacgcc
     6601 tgaatgcgcg tgtgacgtgc cttgctgccc tcgatgcccc gttgcagccc tagatcggcc
     6661 acagcggccg caaacgtggt ctggtcgcgg gtcatctgcg ctttgttgcc gatgaactcc
     6721 ttggccgaca gcctgccgtc ctgcgtcagc ggcaccacga acgcggtcat gtgcgggctg
     6781 gtttcgtcac ggtggatgct ggccgtcacg atgcgatccg ccccgtactt gtccgccagc
     6841 cacttgtgcg ccttctcgaa gaacgccgcc tgctgttctt ggctggccga cttccaccat
     6901 tccgggctgg ccgtcatgac gtactcgacc gccaacacag cgtccttgcg ccgcttctct
     6961 ggcagcaact cgcgcagtcg gcccatcgct tcatcggtgc tgctggccgc ccagtgctcg
     7021 ttctctggcg tcctgctggc gtcagcgttg ggcgtctcgc gctcgcggta ggcgtgcttg
     7081 agactggccg ccacgttgcc cattttcgcc agcttcttgc atcgcatgat cgcgtatgcc
     7141 gccatgcctg cccctccctt ttggtgtcca accggctcga cgggggcagc gcaaggcggt
     7201 gcctccggcg ggccactcaa tgcttgagta tactcactag actttgcttc gcaaagtcgt
     7261 gaccgcctac ggcggctgcg gcgccctacg ggcttgctct ccgggcttcg ccctgcgcgg
     7321 tcgctgcgct cccttgccag cccgtggata tgtggacgat ggccgcga
//
