pET-55-DEST vector is designed to create expression clones (pEXPR) that are fused to a Strep-Tag II coding sequence at the N-terminal.
| 载体基本信息 | |||
| 载体名称 | pET-55-DEST | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 6835 bp | 载体类型 | pET & Duet Vectors (Novagen) |
| 载体来源 | Novagen (EMD Millipore) | 拷贝数 | High copy number |
pET-55-DEST 载体载体图谱
pET-55-DEST 载体载体序列
LOCUS Exported 6835 bp ds-DNA circular SYN 19-11-2013
DEFINITION Gateway(R) Nova bacterial destination vector for expressing proteins
tagged at the N-terminus with Strep-Tag II.
ACCESSION .
VERSION .
KEYWORDS pET-55-DEST
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6835)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6835
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(582..1442)
/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(1443..1535)
/gene="bla"
/note="AmpR promoter"
rep_origin 1616..2204
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2634..2825)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3397..4479)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(4480..4557)
/gene="lacI"
/note="lacI promoter"
/note="
"
promoter 4870..4888
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 4889..4913
/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 4944..4949
/note="ribosome binding site"
CDS 4957..4959
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 4966..4989
/codon_start=1
/product="peptide that binds Strep-Tactin, an engineered
form of streptavidin"
/note="Strep-Tag II"
/translation="WSHPQFEK"
protein_bind 4999..5123
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 5148..5178
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
CDS 5232..5891
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 6233..6538
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(6579..6703)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 6705..6722
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 6763..6810
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
