Basic Vector Information
- Vector Name:
- pOPINM
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6665 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Berrow NS, Alderton D, Sainsbury S, Nettleship J, Assenberg R, Rahman N, Stuart DI, Owens RJ.
- Promoter:
- chicken β-actin
pOPINM 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 ℃.
pOPINM vector Sequence
LOCUS 40924_33792 6665 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pOPINM, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6665)
AUTHORS Berrow NS, Alderton D, Sainsbury S, Nettleship J, Assenberg R,
Rahman N, Stuart DI, Owens RJ.
TITLE A versatile ligation-independent cloning method suitable for
high-throughput expression screening applications
JOURNAL Nucleic Acids Res. 35 (6), E45 (2007)
PUBMED 17317681
REFERENCE 2 (bases 1 to 6665)
AUTHORS Berrow NS.
TITLE Direct Submission
JOURNAL Submitted (17-JAN-2007) Oxford Protein Production Facility, Oxford
University, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, U.K.
REFERENCE 3 (bases 1 to 6665)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6665)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2007"; volume: "35"; issue: "6"; pages: "E45"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-JAN-2007) Oxford Protein Production Facility, Oxford University,
Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, U.K."
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6665
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb 181..1008
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
enhancer 1140..1443
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1449..1726
/label=chicken beta-actin promoter
promoter 2150..2168
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2169..2193
/label=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)."
promoter 2209..2318
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
regulatory 2318..2326
/regulatory_class="ribosome_binding_site"
CDS 2339..2356
/label=6xHis
/note="6xHis affinity tag"
CDS 2366..3466
/label=MBP
/note="maltose binding protein from E. coli"
misc_feature 3480..3494
/label=5' in-fusion site (after KpnI disgest)
/note="5' in-fusion site (after KpnI disgest)"
protein_bind 3540..3561
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3576..3606
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3614..3630
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
CDS 3650..3823
/label=lacZ-alpha
/note="LacZ-alpha fragment of beta-galactosidase"
misc_feature 3829..3846
/label=3' in-fusion site (after HindIII digest)
/note="3' in-fusion site (after HindIII digest)"
CDS 3853..3870
/label=6xHis
/note="6xHis affinity tag"
polyA_signal 4025..4080
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
terminator 4168..4215
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_recomb 4228..4933
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
protein_bind complement(4942..4975)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(5143..5730)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5746..6603)
/label=AmpR
/note="beta-lactamase"
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