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负20摄氏度
6month
pQE- 30 Xa
货源充足
LSM Bio
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2ug
Search name
pQE-30 Xa Information
Promoter: T5
Replicon: ColE1 ori
Terminator: lambda t0 Terminator
Plasmid classification: large intestine plasmid; large intestine expression plasmid; pQE series plasmid.
Plasmid size: 3509bp
Plasmid tagging: N-6 x His, N- Factor Xa site
Prokaryotic resistance: ampicillin Amp (100 g/ml)
Cloning strains: E. coli DH5 and E.
Culture conditions: 37 C, aerobic, LB
Expression host: M15
Culture conditions: 37 C, aerobic, LB
Induction: IPTG or lactose and its analogues.
5'sequencing primers: pQE30-F (TGAGCGGATAACAATTTCAC)
3'sequencing primers: pQE-R (GTTCTGAGGTCATTACTGG)
pQE-30 Xa Description
The pQE-30Xa plasmid contains a powerful T5 promoter (identified by E.coli RNA polymerase) and two lac operon suppression modules, which are used to regulate and express fusion proteins at high level in E.coli. In the presence of Lac inhibitors at high levels, protein synthesis was effectively blocked, and the stability of cytotoxin vector was enhanced. The pQE vector could carry 6 X His tags on the N or C ends of the recombinant protein.
pQE-30 Xa Multiple cloning site
pQE-30 Xa Sequence
LOCUS Exported 3509 bp ds-DNA circular SYN 09-JAN-2013
DEFINITION Bacterial vector for expressing N-terminally 6xHis-tagged proteins
with a Factor Xa cleavage site.
ACCESSION .
VERSION .
KEYWORDS pQE-30 Xa
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3509)
AUTHORS Qiagen
TITLE Direct Submission
JOURNAL Exported Thursday, September 8, 2016 from SnapGene Viewer 3.1.4
COMMENT Clone at StuI to insert a coding sequence just after the Factor Xa
cleavage site.
FEATURES Location/Qualifiers
source 1..3509
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
promoter 10..54
/note="T5 promoter"
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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)."
protein_bind 62..78
/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)."
RBS 101..106
/note="ribosome binding site"
CDS 115..117
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 127..144
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS 166..177
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
misc_feature 175..240
/note="MCS"
/note="multiple cloning site"
misc_feature 240..250
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 256..350
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 394..1053
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1118..1204
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(1685..2273)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2444..3304)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3305..3409)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
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