pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro vector (V010371)

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LOCUS       V010371                11553 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010371
VERSION     V010371
KEYWORDS    pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11553)
  AUTHORS   Cellecta
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 11553)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label="5' LTR (truncated)"
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     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."
     CDS             1496..1540
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1689..1730
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     promoter        1819..2059
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    2065..2094
                     /label="cloning site"
                     /note="cloning site"
                     /note="sgRNA cloning site"
     misc_feature    2122..2237
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
     promoter        2303..2702
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             2721..2741
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2745..2765
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2769..2789
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2796..6896
                     /label="Cas9"
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             6897..6944
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label="bipartite nuclear localization signal from nucl"
                     /note="nucleoplasmin NLS"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6960..7013
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             7020..7616
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    7632..8220
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             8294..8527
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8599..8733
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      8760..8895
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(8976..9006)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(9021..9042)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(9330..9918)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(10092..10949)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(10950..11054)
                     /label="AmpR promoter"
     primer_bind     11528..11544
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"