Basic Vector Information
- Vector Name:
- pOPINVH
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5897 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Nettleship JE, Ren J, Rahman N, Berrow NS, Hatherley D, Barclay AN, Owens RJ.
- Promoter:
- chicken β-actin
pOPINVH 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 ℃.
pOPINVH vector Sequence
LOCUS 40924_33797 5897 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector pOPINVH, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5897) AUTHORS Nettleship JE, Ren J, Rahman N, Berrow NS, Hatherley D, Barclay AN, Owens RJ. TITLE A pipeline for the production of antibody fragments for structural studies using transient expression in HEK 293T cells JOURNAL Protein Expr. Purif. 62 (1), 83-89 (2008) PUBMED 18662785 REFERENCE 2 (bases 1 to 5897) AUTHORS Berrow NS, Owens RJ. TITLE Direct Submission JOURNAL Submitted (20-MAY-2008) Oxford Protein Production Facility, Oxford University, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, UK REFERENCE 3 (bases 1 to 5897) TITLE Direct Submission REFERENCE 4 (bases 1 to 5897) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein Expr. Purif."; date: "2008"; volume: "62"; issue: "1"; pages: "83-89" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (20-MAY-2008) Oxford Protein Production Facility, Oxford University, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, UK" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..5897 /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" misc_feature 2333..2416 /label=mu-phosphatase secretion leader peptide /note="mu-phosphatase secretion leader peptide" misc_feature 2403..2417 /label=5' infusion site after KpnI digest /note="5' infusion site after KpnI digest" protein_bind 2463..2484 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2499..2529 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2537..2553 /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 2573..2746 /label=lacZ-alpha /note="LacZ-alpha fragment of beta-galactosidase" misc_feature 2773..3082 /note="CH1 domain of mouse IgG1 heavy chain; C gamma1" misc_feature 2773..2787 /label=3' infusion site after SfoI digest /note="3' infusion site after SfoI digest" CDS 3085..3102 /label=6xHis /note="6xHis affinity tag" polyA_signal 3257..3312 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" terminator 3400..3447 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_recomb 3460..4165 /label=baculovirus recombination region (ORF1629) /note="contains part of ORF1629" protein_bind complement(4174..4207) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." rep_origin complement(4375..4962) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4978..5835) /label=AmpR /note="beta-lactamase"
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