Basic Vector Information
- Vector Name:
- pOPINE
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5534 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
pOPINE 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 ℃.
pOPINE vector Sequence
LOCUS 40924_33767 5534 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector pOPINE, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5534) 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 5534) 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 5534) TITLE Direct Submission REFERENCE 4 (bases 1 to 5534) 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..5534 /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 2319..2326 /regulatory_class="ribosome_binding_site" misc_feature 2321..2335 /label=5' in-fusion site (after NcoI digest) /note="5' in-fusion site (after NcoI digest)" CDS 2339..2356 /label=6xHis /note="6xHis affinity tag" protein_bind 2409..2430 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2445..2475 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2483..2499 /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 2519..2692 /label=lacZ-alpha /note="LacZ-alpha fragment of beta-galactosidase" CDS 2722..2739 /label=6xHis /note="6xHis affinity tag" polyA_signal 2894..2949 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" terminator 3037..3084 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_recomb 3097..3802 /label=baculovirus recombination region (ORF1629) /note="contains part of ORF1629" protein_bind complement(3811..3844) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." rep_origin complement(4012..4599) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4615..5472) /label=AmpR /note="beta-lactamase"
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