pBBR1MCS- 5 Plasmid

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  • Model: PVT5605
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pBBR1MCS-5

PVT5605 2ug

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pBBR1MCS-5,Plasmid pBBR1MCS-5,pBBR1MCS-5 vector

 

pBBR1MCS-5 Information

Promoter: Lac/lac, T3, T7, or clone promoter

Replicator: pBBR1 Rep, pBBR1 oriV

Plasmid classification: pBBR1MCS series plasmids

Plasmid size: 4768bp

Plasmid label: LacZ

Prokaryotic resistance: gentamicin Gentamycin

Cloned strain: DH5 alpha

Culture conditions: 37 centigrade, aerobic, LB

Host: a wide host

Culture conditions: Reference Literature

5'sequencing primers: M13R:CAGGAAACAGCTATGACC

3'sequencing primers: M13F:TGTAAAACGACGGCCAGT

Use:pBBR5MCS Series plasmid

 

pBBR1MCS-5 Description

pBBR1MCS-5 is constructed based on [1] plasmid pBBR1MCS, the plasmid is constructed by Kovach, has confirmed that [2] can replicate in many Gram-negative bacteria, including Acetobacter xylinum, Alcaligenes eutrophus, Bartonella bacilliformis, whooping cough (Bordetella spp.), by Blue Prand (Brucella spp.), Caulobacter crescentus, Escherichia coli Paracoccous, denitrificans, Pseudomonas fluorescens (Pseudomonas fluorescens), Pseudomonas putida (P. putida), r.meliloti (Rhizobium meliloti), R. leguminosarum by. Viciae, rhodobactersphaeroides (Rhodobacter sphaeroides), Salmonella typhimurium (Salmonella, typhimurium) of Vibrio cholerae (Vibrio cholerae) and Xanthomonas campestris.

Reference

[1] Kovach ME, et al. pBBR1MCS: a broad-host-range cloning vector. Biotechniques, 1994,16(5): 800-802.
[2] Kovach ME, Elzer PH, Hill DS, et al. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene, 1995,166:175-176

pBBR1MCS-5-plasmid

 

 

pBBR1MCS-5 Sequence

LOCUS       Exported                4768 bp ds-DNA     circular SYN 24-JUN-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pBBR1MCS-5

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 4768)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4

FEATURES             Location/Qualifiers

     source          1..4768

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     rep_origin      1023..1792

                     /note="pBBR1 oriV"

                     /note="replication origin of the broad-host-range plasmid 

                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 

                     Rep protein for replication"

     CDS             1793..2455

                     /codon_start=1

                     /product="replication protein for the broad-host-range 

                     plasmid pBBR1 from Bordetella bronchiseptica"

                     /note="pBBR1 Rep"

                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH

                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV

                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE

                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"

     CDS             complement(3013..3378)

                     /codon_start=1

                     /gene="lacZ fragment"

                     /product="LacZ-alpha fragment of beta-galactosidase"

                     /note="lacZ-alpha"

                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV

                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR

                     TDRPSQQLRSLNGEWKL"

     primer_bind     3162..3178

                     /note="M13 fwd"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     promoter        3188..3206

                     /note="T7 promoter"

                     /note="promoter for bacteriophage T7 RNA polymerase"

     misc_feature    3215..3322

                     /note="MCS"

                     /note="pBluescript multiple cloning site"

     primer_bind     3239..3255

                     /note="SK primer"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     primer_bind     complement(3289..3305)

                     /note="KS primer"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     promoter        complement(3335..3353)

                     /note="T3 promoter"

                     /note="promoter for bacteriophage T3 RNA polymerase"

     primer_bind     complement(3374..3390)

                     /note="M13 rev"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     protein_bind    3398..3414

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="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(3422..3452)

                     /note="lac promoter"

                     /note="promoter for the E. coli lac operon"

     protein_bind    3467..3488

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding site"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     promoter        3758..3786

                     /gene="intI1 (promoter lies within the coding sequence)"

                     /note="Pc promoter"

                     /note="class 1 integron promoter"

     CDS             3975..4508

                     /codon_start=1

                     /gene="aacC1"

                     /product="gentamycin acetyltransferase"

                     /note="GmR"

                     /note="confers resistance to gentamycin"

                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD

                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS

                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR

                     EEVMHFDIDPSTAT"

ORIGIN

        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg

       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt

      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg

      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg

      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca

      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt

      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg

      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt

      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg

      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct

      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg

      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc

      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca

      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg

      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg

      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc

      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc

     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga

     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc

     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt

     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg

     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac

     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc

     1381 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg

     1441 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg

     1501 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag

     1561 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta

     1621 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg

     1681 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc

     1741 tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac

     1801 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga

     1861 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct

     1921 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac

     1981 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt

     2041 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt

     2101 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag

     2161 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct

     2221 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga

     2281 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac

     2341 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct

     2401 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt

     2461 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg

     2521 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga

     2581 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga

     2641 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg

     2701 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa

     2761 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa

     2821 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag

     2881 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc

     2941 agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca

     3001 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg

     3061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg

     3121 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga

     3181 gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc

     3241 tagaactagt ggatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga

     3301 cctcgagggg gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct

     3361 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac

     3421 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac

     3481 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc

     3541 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgcatgcata

     3601 aaaactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat

     3661 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg

     3721 acgcacaccg tggaaacgga tgaaggcacg aacccagttg acataagcct gttcggttcg

     3781 taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg

     3841 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttgtaca

     3901 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt

     3961 atggagcagc aacgatgtta cgcagcagca acgatgttac gcagcagggc agtcgcccta

     4021 aaacaaagtt aggtggctca agtatgggca tcattcgcac atgtaggctc ggccctgacc

     4081 aagtcaaatc catgcgggct gctcttgatc ttttcggtcg tgagttcgga gacgtagcca

     4141 cctactccca acatcagccg gactccgatt acctcgggaa cttgctccgt agtaagacat

     4201 tcatcgcgct tgctgccttc gaccaagaag cggttgttgg cgctctcgcg gcttacgttc

     4261 tgcccaggtt tgagcagccg cgtagtgaga tctatatcta tgatctcgca gtctccggcg

     4321 agcaccggag gcagggcatt gccaccgcgc tcatcaatct cctcaagcat gaggccaacg

     4381 cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg tgacgatccc gcagtggctc

     4441 tctatacaaa gttgggcata cgggaagaag tgatgcactt tgatatcgac ccaagtaccg

     4501 ccacctaaca attcgttcaa gccgagatcg gcttcccggc cgcggagttg ttcggtaaat

     4561 tgtcacaacg ccgccaggtg gcacttttcg gggaaatgtg cgcgcccgcg ttcctgctgg

     4621 cgctgggcct gtttctggcg ctggacttcc cgctgttccg tcagcagctt ttcgcccacg

     4681 gccttgatga tcgcggcggc cttggcctgc atatcccgat tcaacggccc cagggcgtcc

     4741 agaacgggct tcaggcgctc ccgaaggt

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