pOPING vector (V004228)

MSDS

Basic Vector Information

Vector Name:: pOPING

Antibiotic Resistance:: Ampicillin

Length:: 5594 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

pOPING vector Vector Map

View Map in NovoBuilder

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 ℃.

pOPING vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_33782        5594 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pOPING, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5594)
  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 5594)
  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 5594)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5594)
  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..5594
                     /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      2333..2425
                     /label=secretion leader peptide (mu -phosphatase)
                     /note="secretion leader peptide (mu -phosphatase)"
                     /regulatory_class="other"
     misc_feature    2409..2423
                     /label=5' in-fusion site (after KpnI digest)
                     /note="5' in-fusion site (after KpnI digest)"
     protein_bind    2469..2490
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2505..2535
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2543..2559
                     /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             2579..2752
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     CDS             2782..2799
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    2954..3009
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      3097..3144
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3157..3862
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(3871..3904)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4072..4659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4675..5532)
                     /label=AmpR
                     /note="beta-lactamase"

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