LOCUS       Exported                9650 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Lentivirus shuttle vector pLV.FLPe.PurR, complete sequence.
ACCESSION   GU253314
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9650)
  AUTHORS   Goncalves MA, Janssen JM, Holkers M, de Vries AA.
  TITLE     Rapid and sensitive lentivirus vector-based conditional gene 
            expression assay to monitor and quantify cell fusion activity
  JOURNAL   PLoS ONE 5 (6), E10954 (2010)
  PUBMED    20532169
REFERENCE   2  (bases 1 to 9650)
  AUTHORS   Goncalves MAF.V., Janssen JM, Holkers M, de Vries AAF.V.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-DEC-2009) Molecular Cell Biology, Leiden University 
            Medical Center, Einthovenweg 20, Leiden, Zuid Holland 2333 ZC, The 
            Netherlands
REFERENCE   3  (bases 1 to 9650)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9650
                     /organism="Lentivirus shuttle vector pLV.FLPe.PurR"
                     /mol_type="other DNA"
                     /note="derived from HIV-1 and FLPe (enhanced version of the
                     Saccharomyces cerevisiae site-specific recombinase FLP)"
                     /db_xref="taxon:715478"
     repeat_region   1..410
                     /note="hybrid 5 LTR derived from Rous sarcoma virus and 
                     HIV-1 LTR sequences"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    454..579
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     regulatory      459..598
                     /regulatory_class="other"
                     /note="HIV-1 packaging signal"
     misc_feature    925..1781
                     /label=Rev-responsive element (RRE)
                     /note="Rev-responsive element (RRE)"
     misc_feature    1072..1305
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1795..1912
                     /label=complementary polypurine tract (cPPT)
                     /note="complementary polypurine tract (cPPT)"
     regulatory      1926..2549
                     /regulatory_class="promoter"
                     /note="immediate-early cytomegalovirus (IE-CMV) gene 
                     promoter"
     enhancer        1935..2238
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2239..2442
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2577..3875
                     /codon_start=1
                     /product="nls/FLPe fusion protein"
                     /label=nls/FLPe fusion protein
                     /note="nls.FLPe; FLPe is an enhanced (thermostable) version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP; fusion protein contains at its N-terminus 
                     corresponding to the SV40 large T antigen nls; 
                     site-specific recombinase"
                     /protein_id="ADC44104.1"
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    4069..4646
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus"
     misc_feature    4069..4641
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4092..4643
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4674..5273
                     /codon_start=1
                     /gene="pac"
                     /product="puromycin N-acetyltransferase"
                     /label=pac
                     /note="puromycin-resistance protein; N-acetyltransferase; 
                     derived from Streptomyces alboniger"
                     /protein_id="ADC44105.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            4674..5273
                     /gene="pac"
                     /label=pac
     misc_feature    5323..6075
                     /note="Homo sapiens hepatitis B virus posttranscriptional 
                     regulatory element"
     repeat_region   6165..6398
                     /label=self-inactivating 3 LTR
                     /note="self-inactivating 3 LTR"
     LTR             6165..6398
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6470..6591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6631..6766
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6787..6805)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6815..6831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6976..7431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7460..7564
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7565..8425
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="prokaryotic ampicillin resistance protein"
                     /protein_id="ADC44106.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8596..9184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9472..9493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9508..9538
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9546..9562
                     /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)."
     primer_bind     9570..9586
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9607..9625
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"