LOCUS       Exported                9413 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pLV.CAG.hrGFP, complete sequence.
ACCESSION   JQ627827
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9413)
  AUTHORS   Pelascini LLP., Janssen JM, Goncalves MAF.V.
  TITLE     Histone deacetylases hinder transgene expression from 
            integration-defective lentiviral vectors in dividing and 
            non-dividing cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9413)
  AUTHORS   Pelascini LLP., Janssen JM, Goncalves MAF.V.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2012) Molecular Cell Biology, Leiden University 
            Medical Center, Einthovenweg 20, Leiden, Zuid Holland 2333 ZC, 
            Netherlands
REFERENCE   3  (bases 1 to 9413)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9413
                     /organism="Shuttle vector pLV.CAG.hrGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1196021"
     repeat_region   1..412
                     /label=RSV-HIV-1 hybrid 5 LTR
                     /note="RSV-HIV-1 hybrid 5 LTR"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     protein_bind    930..1787
                     /label=Rev-responsive element binding site
                     /bound_moiety="Rev-responsive element"
     misc_feature    1077..1310
                     /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    1801..1918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(1945..1992)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      complement(2038..2564)
                     /regulatory_class="polyA_signal_sequence"
                     /note="rabbit beta-globin gene polyadenylation signal"
     polyA_site      2362..2417
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     CDS             complement(2601..3320)
                     /codon_start=1
                     /gene="hrGFP"
                     /product="hrGFP"
                     /label=hrGFP
                     /note="humanized Renilla reniformis green fluorescent 
                     protein"
                     /protein_id="AFL69938.1"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     gene            complement(2601..3320)
                     /gene="hrGFP"
                     /label=hrGFP
     regulatory      complement(3360..5081)
                     /regulatory_class="promoter"
                     /note="CAG chimeric promoter"
     intron          3413..4421
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     promoter        complement(4422..4697)
                     /label=chicken beta-actin promoter
     enhancer        4700..5079
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      5268..5277
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(5330..5377)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      complement(5429..5820)
                     /regulatory_class="polyA_signal_sequence"
                     /note="murine metallothionein gene polyadenylation signal"
     repeat_region   5930..6163
                     /label=HIV-1 SIN 3 LTR
                     /note="HIV-1 SIN 3 LTR"
     LTR             5930..6163
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6235..6356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6396..6531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6552..6570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6580..6596)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6741..7196
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7225..7329
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7330..8190
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AFL69939.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8361..8949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9237..9258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9273..9303
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9311..9327
                     /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     9335..9351
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9372..9390
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"