pBBR1MCS2-pAMP-EGFP Search name
pBBR1MCS2-pAMP-EGFP,Plasmid pBBR1MCS2-pAMP-EGFP,pBBR1MCS2-pAMP-EGFP vector
pBBR1MCS2-pAmp-EGFP Information
Promoter: pAmp
Replicator: pBBR1 Rep, pBBR1 oriV
Plasmid classification: pBBR1MCS series plasmids
Plasmid size: 5996bp
Plasmid label: EGFP
Prokaryotic resistance: kanamycin Kanamycin
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic, LB
Expression host: Gram-negative bacteria
Culture conditions: Reference Literature
5'sequencing primers: M13R:CAGGAAACAGCTATGACC
3'sequencing primers: M13F:TGTAAAACGACGGCCAGT
Remarks: low copy plasmids
Use:pBBR7MCS Series plasmid
pBBR1MCS2-pAmp-EGFP Description
PBBR1MCS2-pAmp-EGFP is based on the construction of plasmid pBBR1MCS, the plasmid is constructed by Kovach, has been shown to replicate in many Gram-negative bacteria, including Acetobacter xylinum, Alcaligenes eutrophus, Bartonella bacilliformis (Bordetella spp.), pertussis, Blue Prand (Brucella spp.), Caulobacter crescentus, Paracoccous denitrificans, Escherichia coli, 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
pBBR1MCS2-pAmp-EGFP Sequence
LOCUS Exported 5996 bp ds-DNA circular SYN 23-JUL-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBBR1MCS2-pAmp-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5996)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, September 10, 2016 from SnapGene Viewer 3.1.4
FEATURES Location/Qualifiers
source 1..5996
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 458..1252
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
protein_bind 1657..1678
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1693..1723
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 1731..1747
/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)."
primer_bind 1755..1771
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
promoter 1792..1810
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 1850..1954
/gene="bla"
/note="AmpR promoter"
CDS 1955..2671
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 2672..2737
/note="MCS"
/note="multiple cloning site"
promoter complement(2791..2809)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2819..2835)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3542..4204)
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pBBR1 from Bordetella bronchiseptica"
/note="pBBR1 Rep"
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
rep_origin 4205..4974
/note="pBBR1 oriV"
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
ORIGIN
1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
181 aaaagtgcca cctgggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
241 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
301 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
361 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
421 aagatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
481 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
541 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
601 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
661 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
721 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
781 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
841 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
901 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
961 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
1021 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
1081 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
1141 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
1201 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
1261 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
1321 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
1381 atcctccagc gcggggatct catgctggag ttcttcgccc acccccatgg gcaaatatta
1441 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
1501 tggcttccat gtcggcagaa tgcttaatga attacaacag <
温馨提示：不可用于临床ZL。
