LOCUS       Exported                7431 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCDH-EF1a-MCS-IRES-zeo, complete sequence.
ACCESSION   KX757249
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7431)
  AUTHORS   Kulemzin SV, Gorchakov AA, Chikaev NA, Volkova OY, Kuznetsova VV, 
            Taranin AV.
  TITLE     Modular lentiviral vector system for optimization of chimeric 
            antigen receptor design
  JOURNAL   Russ. J. Bioorganic Chem. (2016) In press
REFERENCE   2  (bases 1 to 7431)
  AUTHORS   Kulemzin SV, Gorchakov AA, Chikaev NA, Volkova OY, Kuznetsova VV, 
            Taranin AV.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-AUG-2016) Immunogenetics, Institute of Molecular and 
            Cellular Biology, Lavrentyeva 8/2, Novosibirsk 630090, Russia
REFERENCE   3  (bases 1 to 7431)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7431
                     /organism="Cloning vector pCDH-EF1a-MCS-IRES-zeo"
                     /mol_type="other DNA"
                     /db_xref="taxon:1904934"
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     regulatory      7..233
                     /regulatory_class="promoter"
                     /note="hybrid RSV/5LTR promoter"
     LTR             233..413
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from HIV-1"
     repeat_region   234..413
                     /label=HIV LTR
                     /note="HIV LTR"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     regulatory      1807..1924
                     /regulatory_class="enhancer"
                     /note="cPPT"
     misc_feature    1807..1923
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      1936..2479
                     /regulatory_class="promoter"
                     /note="hEF1a-HTLV promoter"
     promoter        1966..2177
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2190..2458
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     misc_feature    2515..3105
                     /label=IRES
                     /note="IRES"
     misc_feature    2524..3110
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     regulatory      3123..3132
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3129..3503
                     /codon_start=1
                     /product="zeocin resistance protein"
                     /label=zeocin resistance protein
                     /protein_id="AOS59239.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    3510..4100
                     /label=WPRE
                     /note="WPRE"
     misc_feature    3510..4098
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3981..3992)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   4172..4405
                     /label=3 deltaLTR
                     /note="3 deltaLTR"
     LTR             4172..4405
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4477..4598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      4638..4773
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4806..4822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4830..4846
                     /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)."
     promoter        complement(4854..4884)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4899..4920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5208..5796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5967..6827)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance protein"
                     /protein_id="AOS59240.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6828..6932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7406..7422
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"