LOCUS       Exported                6916 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pLV.hPGK.eGFP, complete sequence.
ACCESSION   JQ627826
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6916)
  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  JOURNAL   Unpublished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 6916)
  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 6916)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6916
                     /organism="Shuttle vector pLV.hPGK.eGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1196025"
     repeat_region   1..411
                     /label=RSV-HIV-1 hybrid 5 LTR
                     /note="RSV-HIV-1 hybrid 5 LTR"
     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    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     protein_bind    928..1785
                     /label=Rev-responsive element binding site
                     /bound_moiety="Rev-responsive element"
     misc_feature    1075..1308
                     /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    1799..1916
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      2032..2516
                     /regulatory_class="promoter"
                     /note="human phosphoglycerate kinase 1"
     promoter        2033..2516
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     regulatory      2538..2547
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2544..3263
                     /codon_start=1
                     /gene="eGFP"
                     /product="eGFP"
                     /label=eGFP
                     /note="fluorescent reporter"
                     /protein_id="AFL69936.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            2544..3263
                     /gene="eGFP"
                     /label=eGFP
     repeat_region   3430..3663
                     /label=HIV-1 SIN 3 LTR
                     /note="HIV-1 SIN 3 LTR"
     LTR             3430..3663
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3735..3856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3896..4031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(4052..4070)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4080..4096)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4241..4696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4725..4829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4830..5690
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AFL69937.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5861..6449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6737..6758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6774..6804
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6812..6828
                     /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     6836..6852
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6873..6891
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"