PVT10587 2ug
pET303-CT-His Description
pET303-CT-His is an Escherichia coli pET expression plasmid.
Alias: Pet303-ct-his
Promoter: T7/lac promoter
Replicator: ColE1 ori,f1 Ori
Terminator: T7 Terminator
plasmid classification: Escherichia coli carrier; Pet Series Expression plasmid
Plasmid Size: 5369bp
plasmid Label: c-6xhis
prokaryotic Resistance: ampicillin amp
Cloned strain: Escherichia coli dh5α
Culture conditions: 37 鈩? Aerobic, LB
Expression host: Escherichia coli BL21 (DE3)
Culture conditions: 37 鈩? Aerobic, LB
Induction mode: IPTG or lactose and its analogues
5 ' Sequencing primer: T7:TAATACGACTCACTATAGGG 3 ' Sequencing primer: T7-TER:TGCTAGTTATTGCTCAGCGG
The Champion pET302/NT-His and pET303/CT-His Vector Kit is designed for cloning a gene of interest via restriction enzyme(s) and ligase (REaL) and subsequent high-level expression from the strong bacteriophage T7 promoter. In addition to the T7 promoter, each vector contains only the necessary functional elements and an N- or C-terminal 6xHis tag (pET302/NT-His and pET303/CT-His, respectively) for convenient purification and detection (Figure 1). Expression levels obtained from these vectors may be higher than those obtained from another suppliers pET vector (Figure 2). To maximize expression, use with MagicMedia E. coli Expression Medium.
pET303-CT-His Multiple cloning site
pET303-CT-His Sequence
LOCUS Exported 5369 bp ds-DNA circular SYN 23-7-2015
DEFINITION synthetic circular DNA
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5369)
AUTHORS Trial User
TITLE Direct Submission
JOURNAL Exported 2015-7-23
FEATURES Location/Qualifiers
source 1..5369
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 20..38
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 39..63
/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 79..99
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 119..136
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 203..250
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 287..742
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 769..873
/gene="bla"
/note="AmpR promoter"
CDS 874..1734
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1905..2493
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2679..2818
/note="bom"
/note="basis of mobility region from pBR322"
CDS complement(2920..3111)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3920..5002)
/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(5003..5080)
/gene="lacI"
/note="lacI promoter"
ORIGIN
1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
61 tcccctctag taataatttt gtttaacttt aagaaggagg tctagaatgc atctcgagca
121 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
181 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
241 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
301 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
361 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
421 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
481 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
