Basic Vector Information
OpiE2-iFlp-Ieterm 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 ℃.
OpiE2-iFlp-Ieterm vector Sequence
LOCUS 40924_2284 4413 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector OpiE2-iFlp-Ieterm, complete sequence. ACCESSION . 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 TITLE RMCE-based insect cell platform to produce membrane proteins captured on HIV-1 Gag virus-like particles JOURNAL Unpublished REFERENCE 2 (bases 1 to 4413) AUTHORS van den Heuvel JJ, Bleckmann M. TITLE Direct Submission JOURNAL Submitted (05-OCT-2017) Structure and Function of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) TITLE Direct Submission REFERENCE 4 (bases 1 to 4413) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (05-OCT-2017) Structure and Function of Proteins, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, Braunschweig, Lower Saxony 38124, Germany" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Assembly Method :: VectorNTI v. 8 Sequencing Technology :: 454 ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..4413 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 22..569 /label=OpIE-2 promoter /note="strong constitutive baculovirus promoter for insect cell expression" CDS 582..1877 /codon_start=1 /label=FLPo /note="nuclear-targeted site-specific recombinase" /translation="MAPKKKRKVMPQFGILCKTPPKVLVRQFVERFERPSGEKIASCAA ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK LIPAWEFTIIPYYGQKHQSDITDIVSRLQLQFESSEEADKGNSRSKKMLKALLSEGESI WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY QLLKDNLVRSYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI" 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" misc_feature 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) /label=AmpR promoter
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