LOCUS       Exported                8567 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Broad host-range plasmid with toxin-antitoxin system for maintenance
            in the absence of antibiotic selection, and arabinose-inducible 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pBBR5pemIKpBAD
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8567)
  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. 2019 Sep 5;8(36). pii: 8/36/e00769-19. 
            doi: 10.1128/MRA.00769-19.
  PUBMED    31488530
REFERENCE   2  (bases 1 to 8567)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8567
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     161..180
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             265..287
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    302..426
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        451..481
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             535..1194
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(601..620)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1536..1841
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1720..1739
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1882..2006)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2744..2760)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2745..2761)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2790..2810)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2790..2808)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2829..2845)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2829..2845)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2842..2864)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2853..2869
                     /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(2877..2907)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2922..2943
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3213..3241
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3430..3963
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      5245..6016
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             6017..6679
                     /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     7373..7395
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7387..7404
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7388..7404
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7414..7433
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7414..7432
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(7616..8494)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAVNESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        8521..238
                     /gene="araBAD"
                     /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)"
ORIGIN
        1 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
       61 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
      121 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
      181 ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
      241 ccgttttttt gggctagaaa taattttgtt taactttaag aaggagatat acatacccat
      301 cacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata tcaatatatt
      361 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatccagtc
      421 actatggcgg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
      481 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
      541 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
      601 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
      661 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
      721 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
      781 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
      841 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
      901 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
      961 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1021 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     1081 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     1141 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     1201 ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga tttttgcggt
     1261 ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat gctatgaagc
     1321 agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca tatatgatgt
     1381 caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc tgcgtgccga
     1441 acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta ttgaaatgaa
     1501 cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt tacacctata
     1561 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
     1621 ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa gtctcccgtg
     1681 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
     1741 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
     1801 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
     1861 acagccagtc tgcaggtcga ccatagtgac tggatatgtt gtgttttaca gtattatgta
     1921 gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct
     1981 cgttcagctt tcttgtacaa agtggtgatg ggctgcagga attcgatttt cgctataagc
     2041 gtataaatgg aggaattata ttatacttta atataactta acgaaagaag cccatcatgc
     2101 acaccacaaa ccttcgtaaa gtcggtggat cgatcatgct ggccgttccc cccgctttcc
     2161 ttgatcagct tcatcttgag gtcggcgcaa cggtcggctt agccgtcaca gacggtcgtc
     2221 tggtcattga gccagtccta catcctcagt acacgctgga tcaactgctg gccgaagcgg
     2281 aagcctccgg ggcttacccg ttgccgcccg aggaacgtga atgggtcgat gctccagcag
     2341 taggacgcga actgatatga aacgcggcga tgtttacatg gtcgatctgg agccaaccgc
     2401 cggacacgaa caacgcggtc accgtccggt ggttgttatc tccagtgagc gattcaatcg
     2461 gcttacctct tgtccggtga tccttccaat cacaaacggc ggcgagttcg cctcccgcct
     2521 cgggttcgct gtggaattac taggtacgat tacaacgggc gtcgttcgat gtgatcaacc
     2581 acgtgcgctc gacttactcg ctcgcaatgc ccgcaaagtg gaaagcctac caccaaacat
     2641 cttggccgat gtcttggcaa aagctgtcac catcttccag taatttttgg ggcaagttca
     2701 ggccactgca accaatcact agtgaattcg atatcaagct tatcgatacc gtcgacctcg
     2761 agggggggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     2821 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2881 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2941 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3001 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcat gcataaaaac
     3061 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     3121 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggacgca
     3181 caccgtggaa acggatgaag gcacgaaccc agttgacata agcctgttcg gttcgtaaac
     3241 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     3301 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta
     3361 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     3421 gcagcaacga tgttacgcag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
     3481 aagttaggtg gctcaagtat gggcatcatt cgcacatgta ggctcggccc tgaccaagtc
     3541 aaatccatgc gggctgctct tgatcttttc ggtcgtgagt tcggagacgt agccacctac
     3601 tcccaacatc agccggactc cgattacctc gggaacttgc tccgtagtaa gacattcatc
     3661 gcgcttgctg ccttcgacca agaagcggtt gttggcgctc tcgcggctta cgttctgccc
     3721 aagtttgagc agccgcgtag tgagatctat atctatgatc tcgcagtctc cggagagcac
     3781 cggaggcagg gcattgccac cgcgctcatc aatctcctca agcatgaggc caacgcgctt
     3841 ggtgcttatg tgatctacgt gcaagcagat tacggtgacg atcccgcagt ggctctctat
     3901 acaaagttgg gcatacggga agaagtgatg cactttgata tcgacccaag taccgccacc
     3961 taacaattcg ttcaagccga gatcggcttc ccggccgcgg agttgttcgg taaattgtca
     4021 caacgccgca ggtggcactt ttcggggaaa tgtgcgcgcc cgcgttcctg ctggcgctgg
     4081 gcctgtttct ggcgctggac ttcccgctgt tccgtcagca gcttttcgcc cacggccttg
     4141 atgatcgcgg cggccttggc ctgcatatcc cgattcaacg gccccagggc gtccagaacg
     4201 ggcttcaggc gctcccgaag gtctcgggcc gtctcttggg cttgatcggc cttcttgcgc
     4261 atctcacgcg ctcctgcggc ggcctgtagg gcaggctcat acccctgccg aaccgctttt
     4321 gtcagccggt cggccacggc ttccggcgtc tcaacgcgct ttgagattcc cagcttttcg
     4381 gccaatccct gcggtgcata ggcgcgtggc tcgaccgctt gcgggctgat ggtgacgtgg
     4441 cccactggtg gccgctccag ggcctcgtag aacgcctgaa tgcgcgtgtg acgtgccttg
     4501 ctgccctcga tgccccgttg cagccctaga tcggccacag cggccgcaaa cgtggtctgg
     4561 tcgcgggtca tctgcgcttt gttgccgatg aactccttgg ccgacagcct gccgtcctgc
     4621 gtcagcggca ccacgaacgc ggtcatgtgc gggctggttt cgtcacggtg gatgctggcc
     4681 gtcacgatgc gatccgcccc gtacttgtcc gccagccact tgtgcgcctt ctcgaagaac
     4741 gccgcctgct gttcttggct ggccgacttc caccattccg ggctggccgt catgacgtac
     4801 tcgaccgcca acacagcgtc cttgcgccgc ttctctggca gcaactcgcg cagtcggccc
     4861 atcgcttcat cggtgctgct ggccgcccag tgctcgttct ctggcgtcct gctggcgtca
     4921 gcgttgggcg tctcgcgctc gcggtaggcg tgcttgagac tggccgccac gttgcccatt
     4981 ttcgccagct tcttgcatcg catgatcgcg tatgccgcca tgcctgcccc tcccttttgg
     5041 tgtccaaccg gctcgacggg ggcagcgcaa ggcggtgcct ccggcgggcc actcaatgct
     5101 tgagtatact cactagactt tgcttcgcaa agtcgtgacc gcctacggcg gctgcggcgc
     5161 cctacgggct tgctctccgg gcttcgccct gcgcggtcgc tgcgctccct tgccagcccg
     5221 tggatatgtg gacgatggcc gcgagcggcc accggctggc tcgcttcgct cggcccgtgg
     5281 acaaccctgc tggacaagct gatggacagg ctgcgcctgc ccacgagctt gaccacaggg
     5341 attgcccacc ggctacccag ccttcgacca catacccacc ggctccaact gcgcggcctg
     5401 cggccttgcc ccatcaattt ttttaatttt ctctggggaa aagcctccgg cctgcggcct
     5461 gcgcgcttcg cttgccggtt ggacaccaag tggaaggcgg gtcaaggctc gcgcagcgac
     5521 cgcgcagcgg cttggccttg acgcgcctgg aacgacccaa gcctatgcga gtgggggcag
     5581 tcgaagggcg aagcccgccc gcctgccccc cgagcctcac ggcggcgagt gcgggggttc
     5641 caagggggca gcgccacctt gggcaaggcc gaaggccgcg cagtcgatca acaagccccg
     5701 gaggggccac tttttgccgg agggggagcc gcgccgaagg cgtgggggaa ccccgcaggg
     5761 gtgcccttct ttgggcacca aagaactaga tatagggcga aatgcgaaag acttaaaaat
     5821 caacaactta aaaaaggggg gtacgcaaca gctcattgcg gcaccccccg caatagctca
     5881 ttgcgtaggt taaagaaaat ctgtaattga ctgccacttt tacgcaacgc ataattgttg
     5941 tcgcgctgcc gaaaagttgc agctgattgc gcatggtgcc gcaaccgtgc ggcaccccta
     6001 ccgcatggag ataagcatgg ccacgcagtc cagagaaatc ggcattcaag ccaagaacaa
     6061 gcccggtcac tgggtgcaaa cggaacgcaa agcgcatgag gcgtgggccg ggcttattgc
     6121 gaggaaaccc acggcggcaa tgctgctgca tcacctcgtg gcgcagatgg gccaccagaa
     6181 cgccgtggtg gtcagccaga agacactttc caagctcatc ggacgttctt tgcggacggt
     6241 ccaatacgca gtcaaggact tggtggccga gcgctggatc tccgtcgtga agctcaacgg
     6301 ccccggcacc gtgtcggcct acgtggtcaa tgaccgcgtg gcgtggggcc agccccgcga
     6361 ccagttgcgc ctgtcggtgt tcagtgccgc cgtggtggtt gatcacgacg accaggacga
     6421 atcgctgttg gggcatggcg acctgcgccg catcccgacc ctgtatccgg gcgagcagca
     6481 actaccgacc ggccccggcg aggagccgcc cagccagccc ggcattccgg gcatggaacc
     6541 agacctgcca gccttgaccg aaacggagga atgggaacgg cgcgggcagc agcgcctgcc
     6601 gatgcccgat gagccgtgtt ttctggacga tggcgagccg ttggagccgc cgacacgggt
     6661 cacgctgccg cgccggtagc acttgggttg cgcagcaacc cgtaagtgcg ctgttccaga
     6721 ctatcggctg tagccgcctc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
     6781 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
     6841 caccgttttt atcaggctct gggaggcaga ataaatgatc atatcgtcaa ttattacctc
     6901 cacggggaga gcctgagcaa actggcctca ggcatttgag aagcacacgg tcacactgct
     6961 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
     7021 ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta
     7081 tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc ttaaaaaaat
     7141 tacgccccgc cctgccactc atcgcagtac ggcctattgg ttaaaaaatg agctgattta
     7201 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
     7261 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     7321 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     7381 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     7441 gagctcgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca
     7501 attcgcgcgc gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga
     7561 ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg
     7621 acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc
     7681 tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat
     7741 tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga
     7801 tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg
     7861 acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt
     7921 ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat
     7981 ggagcgactc gttaaccgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg
     8041 ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt
     8101 cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg
     8161 acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat
     8221 accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca
     8281 aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc
     8341 gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga
     8401 gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc
     8461 ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag
     8521 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtc
//
