LOCUS       Exported                7894 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCDH-EF1a-mIgkSP-MCS-IRES-copGFP, complete sequence.
ACCESSION   KX757241
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7894)
  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 7894)
  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 7894)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7894
                     /organism="Cloning vector
                     pCDH-EF1a-mIgkSP-MCS-IRES-copGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1904942"
     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"
     regulatory      2484..2493
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2490..2552
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     misc_feature    2495..2548
                     /label=mouse Igk signal peptide
                     /note="mouse Igk signal peptide"
     misc_feature    2588..3178
                     /label=IRES
                     /note="IRES"
     misc_feature    2597..3183
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3196..3954
                     /codon_start=1
                     /product="copGFP"
                     /label=copGFP
                     /note="green fluorescent protein from copepod Pontellina 
                     plumata"
                     /protein_id="AOS59217.1"
                     /translation="MESDESGLPAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKM
                     KSTKGALTFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVL
                     HVSFSYRYEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFAR
                     TFSLRDGGYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAF
                     KTPIAFARSRAQSSNSAVDGTAGPGSTGSR"
     CDS             3220..3882
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     misc_feature    3973..4563
                     /label=WPRE
                     /note="WPRE"
     misc_feature    3973..4561
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4444..4455)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   4635..4868
                     /label=3 deltaLTR
                     /note="3 deltaLTR"
     LTR             4635..4868
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4940..5061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5101..5236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5269..5285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5293..5309
                     /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(5317..5347)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5362..5383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5671..6259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6430..7290)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /note="ampicillin resistance protein"
                     /protein_id="AOS59218.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6430..7290)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(7291..7395)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7869..7885
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"