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  JOURNAL   Unpublished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