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OpiE2-iFlp-Ieterm vector (V009821#)

Basic Vector Information

Basic Vector Information
Vector Name OpiE2-iFlp-Ieterm Antibiotic Resistance Ampicillin
Length 4413 bp Type Cloning vector
Source Vidigal J, Teixeira AP.

OpiE2-iFlp-Ieterm vector Vector Map

iFLP ori beta-lactamase AmpR promoter

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 ℃.

OpiE2-iFlp-Ieterm vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                4413 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector OpiE2-iFlp-Ieterm, complete sequence.
ACCESSION   MG051710
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4413)
  AUTHORS   Vidigal J, Teixeira AP.
  TITLE     RMCE-based insect cell platform to produce membrane proteins 
            captured on HIV-1 Gag virus-like particles
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4413)
  AUTHORS   van den Heuvel JJ, Bleckmann M.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-OCT-2017) Structure and Function of Proteins, 
            Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 
            Braunschweig, Lower Saxony 38124, Germany
REFERENCE   3  (bases 1 to 4413)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: VectorNTI v. 8
            Sequencing Technology :: 454
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4413
                     /organism="Cloning vector OpiE2-iFlp-Ieterm"
                     /mol_type="other DNA"
                     /db_xref="taxon:2049880"
     regulatory      21..573
                     /regulatory_class="promoter"
     promoter        22..569
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect 
                     cell expression"
     CDS             582..1886
                     /codon_start=1
                     /product="iFLP"
                     /label=iFLP
                     /note="optimized flippase for expression in insect cells 
                     Sf21"
                     /protein_id="ATP16879.1"
                     /translation="MAPKKKRKVMPQFGILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYYGQKHQSDITDIVSRLQLQFESSEEADKGNSRSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRIVD"
     CDS             588..608
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1893..2199
                     /regulatory_class="terminator"
     terminator      1893..2199
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     rep_origin      complement(2553..3141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2622..2903
     CDS             complement(3535..4170)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance protein"
                     /protein_id="ATP16880.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRTINWRTTYSSFPATINRLDGGG"
     CDS             complement(3608..3986)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     promoter        complement(4171..4275)
                     /gene="bla"
                     /label=AmpR promoter

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