LOCUS       sequence-245912-        7189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host range plasmid derived from pBBR1-MCS5 with pemI/pemK 
            toxin-antitoxin module from Xylella fastidiosa to improve stability 
            in the absence of antibiotic selection, Gateway cloning compatible.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7189)
  AUTHORS   Burbank LP, Stenger DC
  TITLE     Plasmid Vectors for Xylella fastidiosa Utilizing a Toxin-Antitoxin 
            System for Stability in the Absence of Antibiotic Selection.
  JOURNAL   Phytopathology. 2016 Aug;106(8):928-36. doi: 
            10.1094/PHYTO-02-16-0097-R. Epub 2016 Jun 7.
  PUBMED    27088393
REFERENCE   2  (bases 1 to 7189)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1094/PHYTO-02-16-0097-R"; journalName: "Phytopathology"; date: 
            "2016-08"; volume: "106"; issue: "8"; pages: "928-36"
COMMENT     Sequence Label: pBBR5pemIK-GW
FEATURES             Location/Qualifiers
     source          1..7189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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     1894..1916
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1908..1925
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1909..1925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1935..1954
                     /label=T7
                     /note="T7 promoter, forward primer"
     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"
     primer_bind     1986..2002
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2011..2135
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2160..2165,2166..2183,2184..2190)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2244..2903
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2310..2329)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3245..3550
                     /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     3429..3448
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3591..3715)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4454..4470)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4455..4471)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4500..4520)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4500..4518)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4539..4555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4539..4555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4552..4574)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4563..4579
                     /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(join(4587..4593,4594..4611,4612..4617))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4632..4653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4923..4928,4929..4945,4946..4951)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5140..5673
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      join(6955..7189,1..537)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 tggacaccaa gtggaaggcg ggtcaaggct cgcgcagcga ccgcgcagcg gcttggcctt
       61 gacgcgcctg gaacgaccca agcctatgcg agtgggggca gtcgaagggc gaagcccgcc
      121 cgcctgcccc ccgagcctca cggcggcgag tgcgggggtt ccaagggggc agcgccacct
      181 tgggcaaggc cgaaggccgc gcagtcgatc aacaagcccc ggaggggcca ctttttgccg
      241 gagggggagc cgcgccgaag gcgtggggga accccgcagg ggtgcccttc tttgggcacc
      301 aaagaactag atatagggcg aaatgcgaaa gacttaaaaa tcaacaactt aaaaaagggg
      361 ggtacgcaac agctcattgc ggcacccccc gcaatagctc attgcgtagg ttaaagaaaa
      421 tctgtaattg actgccactt ttacgcaacg cataattgtt gtcgcgctgc cgaaaagttg
      481 cagctgattg cgcatggtgc cgcaaccgtg cggcacccct accgcatgga gataagcatg
      541 gccacgcagt ccagagaaat cggcattcaa gccaagaaca agcccggtca ctgggtgcaa
      601 acggaacgca aagcgcatga ggcgtgggcc gggcttattg cgaggaaacc cacggcggca
      661 atgctgctgc atcacctcgt ggcgcagatg ggccaccaga acgccgtggt ggtcagccag
      721 aagacacttt ccaagctcat cggacgttct ttgcggacgg tccaatacgc agtcaaggac
      781 ttggtggccg agcgctggat ctccgtcgtg aagctcaacg gccccggcac cgtgtcggcc
      841 tacgtggtca atgaccgcgt ggcgtggggc cagccccgcg accagttgcg cctgtcggtg
      901 ttcagtgccg ccgtggtggt tgatcacgac gaccaggacg aatcgctgtt ggggcatggc
      961 gacctgcgcc gcatcccgac cctgtatccg ggcgagcagc aactaccgac cggccccggc
     1021 gaggagccgc ccagccagcc cggcattccg ggcatggaac cagacctgcc agccttgacc
     1081 gaaacggagg aatgggaacg gcgcgggcag cagcgcctgc cgatgcccga tgagccgtgt
     1141 tttctggacg atggcgagcc gttggagccg ccgacacggg tcacgctgcc gcgccggtag
     1201 cacttgggtt gcgcagcaac ccgtaagtgc gctgttccag actatcggct gtagccgcct
     1261 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     1321 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccgtttt tatcaggctc
     1381 tgggaggcag aataaatgat catatcgtca attattacct ccacggggag agcctgagca
     1441 aactggcctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     1501 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     1561 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     1621 aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
     1681 catcgcagta cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1741 tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg
     1801 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     1861 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     1921 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctccac cgcggtggcg
     1981 gccgctctag aactagtgga tcccccatca acaagtttgt acaaaaaagc tgaacgagaa
     2041 acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa acagactaca
     2101 taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct
     2161 ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat
     2221 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     2281 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     2341 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     2401 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     2461 tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     2521 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     2581 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     2641 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     2701 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     2761 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     2821 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     2881 atgagtggca gggcggggcg taaagatctg gatccggctt actaaaagcc agataacagt
     2941 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     3001 gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag
     3061 ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg
     3121 cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg
     3181 gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg
     3241 tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     3301 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
     3361 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     3421 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     3481 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     3541 gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact
     3601 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     3661 tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggttgatg
     3721 ggctgcagga attcgatttt cgctataagc gtataaatgg aggaattata ttatacttta
     3781 atataactta acgaaagaag cccatcatgc acaccacaaa ccttcgtaaa gtcggtggat
     3841 cgatcatgct ggccgttccc cccgctttcc ttgatcagct tcatcttgag gtcggcgcaa
     3901 cggtcggctt agccgtcaca gacggtcgtc tggtcattga gccagtccta catcctcagt
     3961 acacgctgga tcaactgctg gccgaagcgg aagcctccgg ggcttacccg ttgccgcccg
     4021 aggaacgtga atgggtcgat gctccagcag taggacgcga actgatatga aacgcggcga
     4081 tgtttacatg gtcgatctgg agccaaccgc cggacacgaa caacgcggtc accgtccggt
     4141 ggttgttatc tccagtgagc gattcaatcg gcttacctct tgtccggtga tccttccaat
     4201 cacaaacggc ggcgagttcg cctcccgcct cgggttcgct gtggaattac taggtacgat
     4261 tacaacgggc gtcgttcgat gtgatcaacc acgtgcgctc gacttactcg ctcgcaatgc
     4321 ccgcaaagtg gaaagcctac caccaaacat cttggccgat gtcttggcaa aagctgtcac
     4381 catcttccag taatttttgg ggcaagttca ggccactgca accaatcact agtgaattcg
     4441 atatcaagct tatcgatacc gtcgacctcg agggggggcc cggtacccag cttttgttcc
     4501 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     4561 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4621 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4681 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4741 ggtttgcgta ttgggcgcat gcataaaaac tgttgtaatt cattaagcat tctgccgaca
     4801 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     4861 ccttgcgtat aatatttgcc catggacgca caccgtggaa acggatgaag gcacgaaccc
     4921 agttgacata agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca
     4981 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     5041 cttgttatga ctgttttttt gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     5101 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagcaacgat
     5161 gttacgcagc agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt
     5221 cgcacatgta ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc
     5281 ggtcgtgagt tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc
     5341 gggaacttgc tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt
     5401 gttggcgctc tcgcggctta cgttctgccc aagtttgagc agccgcgtag tgagatctat
     5461 atctatgatc tcgcagtctc cggagagcac cggaggcagg gcattgccac cgcgctcatc
     5521 aatctcctca agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat
     5581 tacggtgacg atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg
     5641 cactttgata tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc
     5701 ccggccgcgg agttgttcgg taaattgtca caacgccgca ggtggcactt ttcggggaaa
     5761 tgtgcgcgcc cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac ttcccgctgt
     5821 tccgtcagca gcttttcgcc cacggccttg atgatcgcgg cggccttggc ctgcatatcc
     5881 cgattcaacg gccccagggc gtccagaacg ggcttcaggc gctcccgaag gtctcgggcc
     5941 gtctcttggg cttgatcggc cttcttgcgc atctcacgcg ctcctgcggc ggcctgtagg
     6001 gcaggctcat acccctgccg aaccgctttt gtcagccggt cggccacggc ttccggcgtc
     6061 tcaacgcgct ttgagattcc cagcttttcg gccaatccct gcggtgcata ggcgcgtggc
     6121 tcgaccgctt gcgggctgat ggtgacgtgg cccactggtg gccgctccag ggcctcgtag
     6181 aacgcctgaa tgcgcgtgtg acgtgccttg ctgccctcga tgccccgttg cagccctaga
     6241 tcggccacag cggccgcaaa cgtggtctgg tcgcgggtca tctgcgcttt gttgccgatg
     6301 aactccttgg ccgacagcct gccgtcctgc gtcagcggca ccacgaacgc ggtcatgtgc
     6361 gggctggttt cgtcacggtg gatgctggcc gtcacgatgc gatccgcccc gtacttgtcc
     6421 gccagccact tgtgcgcctt ctcgaagaac gccgcctgct gttcttggct ggccgacttc
     6481 caccattccg ggctggccgt catgacgtac tcgaccgcca acacagcgtc cttgcgccgc
     6541 ttctctggca gcaactcgcg cagtcggccc atcgcttcat cggtgctgct ggccgcccag
     6601 tgctcgttct ctggcgtcct gctggcgtca gcgttgggcg tctcgcgctc gcggtaggcg
     6661 tgcttgagac tggccgccac gttgcccatt ttcgccagct tcttgcatcg catgatcgcg
     6721 tatgccgcca tgcctgcccc tcccttttgg tgtccaaccg gctcgacggg ggcagcgcaa
     6781 ggcggtgcct ccggcgggcc actcaatgct tgagtatact cactagactt tgcttcgcaa
     6841 agtcgtgacc gcctacggcg gctgcggcgc cctacgggct tgctctccgg gcttcgccct
     6901 gcgcggtcgc tgcgctccct tgccagcccg tggatatgtg gacgatggcc gcgagcggcc
     6961 accggctggc tcgcttcgct cggcccgtgg acaaccctgc tggacaagct gatggacagg
     7021 ctgcgcctgc ccacgagctt gaccacaggg attgcccacc ggctacccag ccttcgacca
     7081 catacccacc ggctccaact gcgcggcctg cggccttgcc ccatcaattt ttttaatttt
     7141 ctctggggaa aagcctccgg cctgcggcct gcgcgcttcg cttgccggt
//
