LOCUS       62056_3740        8590 bp DNA     circular SYN 06-DEC-2019
DEFINITION  Cloning vector pBBR5pemIK-pBAD, complete sequence.
ACCESSION   MN044104
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8590)
  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 8590)
  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 8590)
  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..8590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..772
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             773..1432
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     primer_bind     2144..2160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2170..2188
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2375..3250)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        3277..3561
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     RBS             3588..3610
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     3622..3638
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3647..3771
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3796..3826
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             3880..4536
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             4881..5183
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(5227..5351)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5443..5706
                     /codon_start=1
                     /transl_table=11
                     /product="antitoxin PemI"
                     /label=antitoxin PemI
                     /protein_id="QDJ95343.1"
                     /translation="MHTTNLRKVGGSIMLAVPPAFLDQLHLEVGATVGLAVTDGRLVIE
                     PVLHPQYTLDQLLAEAEASGAYPLPPEEREWVDAPAVGRELI"
     CDS             5667..6029
                     /codon_start=1
                     /transl_table=11
                     /product="mRNA interferase PemK"
                     /label=mRNA interferase PemK
                     /protein_id="QDJ95344.1"
                     /translation="MGRCSSSRTRTDMKRGDVYMVDLEPTAGHEQRGHRPVVVISSERF
                     NRLTSCPVILPITNGGEFASRLGFAVELLGTITTGVVRCDQPRALDLLARNARKVESLP
                     PNILADVLAKAVTIFQ"
     primer_bind     complement(6090..6106)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6136..6154)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6175..6191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6199..6215)
                     /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(6223..6253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6268..6289)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6559..6587
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6776..7306
                     /label=GmR
                     /note="gentamycin acetyltransferase"
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 aaggcgaagc ccgcccgcct
      361 gccccccgag cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc caccttgggc
      421 aaggccgaag gccgcgcagt cgatcaacaa gccccggagg ggccactttt tgccggaggg
      481 ggagccgcgc cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg gcaccaaaga
      541 actagatata gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa aggggggtac
      601 gcaacagctc attgcggcac cccccgcaat agctcattgc gtaggttaaa gaaaatctgt
      661 aattgactgc cacttttacg caacgcataa ttgttgtcgc gctgccgaaa agttgcagct
      721 gattgcgcat ggtgccgcaa ccgtgcggca cccctaccgc atggagataa gcatggccac
      781 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
      841 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
      901 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
      961 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     1021 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     1081 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     1141 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     1201 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     1261 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     1321 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     1381 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     1441 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     1501 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     1561 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga
     1621 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg
     1681 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     1741 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     1801 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca
     1861 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     1921 cagtacggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     1981 caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg
     2041 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     2101 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     2161 gagcgcgcgt aatacgactc actatagggc gaattggagc tcgcatgtgt cagaggtttt
     2221 caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc
     2281 atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg
     2341 tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca
     2401 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     2461 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcggtaggca
     2521 tccgggtggt gctcaaaagc agcttcgcct ggctggtacg ttggtcctcg cgccagctta
     2581 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     2641 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     2701 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     2761 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     2821 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     2881 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     2941 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     3001 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcagacaa
     3061 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     3121 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     3181 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     3241 cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca
     3301 tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc
     3361 cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca
     3421 aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt
     3481 gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc
     3541 aactctctac tgtttctcca tacccgtttt tttgggctag aaataatttt gtttaacttt
     3601 aagaaggaga tatacatacc ctctagaact agtggatccc ccatcaacaa gtttgtacaa
     3661 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
     3721 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca
     3781 ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt
     3841 taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     3901 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     3961 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     4021 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     4081 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
     4141 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     4201 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     4261 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     4321 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     4381 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     4441 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     4501 ttacaacagt actgcgatga gtggcagggc ggggcgtaaa gatctggatc cggcttacta
     4561 aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact
     4621 gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga
     4681 cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct
     4741 ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga
     4801 aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga
     4861 cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt
     4921 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     4981 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     5041 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     5101 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     5161 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag
     5221 gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     5281 aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg
     5341 tacaaagtgg ttgatgggct gcaggaattc gattttcgct ataagcgtat aaatggagga
     5401 attatattat actttaatat aacttaacga aagaagccca tcatgcacac cacaaacctt
     5461 cgtaaagtcg gtggatcgat catgctggcc gttccccccg ctttccttga tcagcttcat
     5521 cttgaggtcg gcgcaacggt cggcttagcc gtcacagacg gtcgtctggt cattgagcca
     5581 gtcctacatc ctcagtacac gctggatcaa ctgctggccg aagcggaagc ctccggggct
     5641 tacccgttgc cgcccgagga acgtgaatgg gtcgatgctc cagcagtagg acgcgaactg
     5701 atatgaaacg cggcgatgtt tacatggtcg atctggagcc aaccgccgga cacgaacaac
     5761 gcggtcaccg tccggtggtt gttatctcca gtgagcgatt caatcggctt acctcttgtc
     5821 cggtgatcct tccaatcaca aacggcggcg agttcgcctc ccgcctcggg ttcgctgtgg
     5881 aattactagg tacgattaca acgggcgtcg ttcgatgtga tcaaccacgt gcgctcgact
     5941 tactcgctcg caatgcccgc aaagtggaaa gcctaccacc aaacatcttg gccgatgtct
     6001 tggcaaaagc tgtcaccatc ttccagtaat ttttggggca agttcaggcc actgcaacca
     6061 atcactagtg aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt
     6121 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     6181 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6241 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6301 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6361 acgcgcgggg agaggcggtt tgcgtattgg gcgcatgcat aaaaactgtt gtaattcatt
     6421 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     6481 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gacgcacacc gtggaaacgg
     6541 atgaaggcac gaacccagtt gacataagcc tgttcggttc gtaaactgta atgcaagtag
     6601 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     6661 gtggcggttt tcatggcttg ttatgactgt ttttttgtac agtctatgcc tcgggcatcc
     6721 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     6781 acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggtggctc
     6841 aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat ccatgcgggc
     6901 tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc aacatcagcc
     6961 ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt
     7021 cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc
     7081 gcgtagtgag atctatatct atgatctcgc agtctccgga gagcaccgga ggcagggcat
     7141 tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg cttatgtgat
     7201 ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa agttgggcat
     7261 acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcgttca
     7321 agccgagatc ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac gccgcaggtg
     7381 gcacttttcg gggaaatgtg cgcgcccgcg ttcctgctgg cgctgggcct gtttctggcg
     7441 ctggacttcc cgctgttccg tcagcagctt ttcgcccacg gccttgatga tcgcggcggc
     7501 cttggcctgc atatcccgat tcaacggccc cagggcgtcc agaacgggct tcaggcgctc
     7561 ccgaaggtct cgggccgtct cttgggcttg atcggccttc ttgcgcatct cacgcgctcc
     7621 tgcggcggcc tgtagggcag gctcataccc ctgccgaacc gcttttgtca gccggtcggc
     7681 cacggcttcc ggcgtctcaa cgcgctttga gattcccagc ttttcggcca atccctgcgg
     7741 tgcataggcg cgtggctcga ccgcttgcgg gctgatggtg acgtggccca ctggtggccg
     7801 ctccagggcc tcgtagaacg cctgaatgcg cgtgtgacgt gccttgctgc cctcgatgcc
     7861 ccgttgcagc cctagatcgg ccacagcggc cgcaaacgtg gtctggtcgc gggtcatctg
     7921 cgctttgttg ccgatgaact ccttggccga cagcctgccg tcctgcgtca gcggcaccac
     7981 gaacgcggtc atgtgcgggc tggtttcgtc acggtggatg ctggccgtca cgatgcgatc
     8041 cgccccgtac ttgtccgcca gccacttgtg cgccttctcg aagaacgccg cctgctgttc
     8101 ttggctggcc gacttccacc attccgggct ggccgtcatg acgtactcga ccgccaacac
     8161 agcgtccttg cgccgcttct ctggcagcaa ctcgcgcagt cggcccatcg cttcatcggt
     8221 gctgctggcc gcccagtgct cgttctctgg cgtcctgctg gcgtcagcgt tgggcgtctc
     8281 gcgctcgcgg taggcgtgct tgagactggc cgccacgttg cccattttcg ccagcttctt
     8341 gcatcgcatg atcgcgtatg ccgccatgcc tgcccctccc ttttggtgtc caaccggctc
     8401 gacgggggca gcgcaaggcg gtgcctccgg cgggccactc aatgcttgag tatactcact
     8461 agactttgct tcgcaaagtc gtgaccgcct acggcggctg cggcgcccta cgggcttgct
     8521 ctccgggctt cgccctgcgc ggtcgctgcg ctcccttgcc agcccgtgga tatgtggacg
     8581 atggccgcga
//
