Basic Vector Information
red fluorescent protein expression plasmid; wide host plasmid.
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pBBR1MCS2-Tac-mCherry | Antibiotic Resistance | Kanamycin |
| Length | 6195 bp | Type | Expression plasmid |
| Promoter | Tac | Cloning Method | Enzyme Cut |
| 5' Primer | M13F:TGTAAAACGACGGCCAGT | 3' Primer | M13R:CAGGAAACAGCTATGACC |
| Fusion Tag | mCherry | ||
pBBR1MCS2-Tac-mCherry vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pBBR1MCS2-Tac-mCherry vector Sequence
LOCUS Exported 6195 bp ds-DNA circular SYN 15-JUL-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBBR1MCS2-Tac-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6195)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6195
/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"
primer_bind 1840..1856
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(1910..1987)
/gene="lacI"
/note="lacI promoter"
misc_feature 2033..2061
/note="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
CDS 2108..2818
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/note="mCherry"
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
terminator 2852..2899
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(2941..2957)
/note="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2990..3008)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3018..3034)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(3741..4403)
/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 4404..5173
/note="pBBR1 oriV"
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
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