pRSI17cb-U6-sh-13kCB18-HTS6-UbiC-TagGFP2-2A-Puro vector (V010366)

LOCUS       V010366                 8050 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010366
VERSION     V010366
KEYWORDS    pRSI17cb-U6-sh-13kCB18-HTS6-UbiC-TagGFP2-2A-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8050)
  AUTHORS   Cellecta
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8050)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8050
                     /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="shRNA cloning site"
     gap             2095..2112
                     /estimated_length=18
     misc_feature    2095..2112
                     /label="clonal barcode"
                     /note="clonal barcode"
     misc_feature    2144..2259
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
     promoter        2325..2724
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     CDS             2737..3450
                     /label="TagGFP2"
                     /note="monomeric green fluorescent protein, also known as
                     mTagGFP"
     CDS             3457..3510
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label="2A peptide from Thosea asigna virus capsid protein"
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3517..4113
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4129..4717
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4791..5024
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5096..5230
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5257..5392
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5425..5441)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5449..5465)
                     /label="lac operator"
                     /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(5473..5503)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5518..5539)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5827..6415)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6589..7446)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7447..7551)
                     /label="AmpR promoter"
     primer_bind     8025..8041
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"