CRISPRi library backbone vector (V012348)

LOCUS       V012348                 8888 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V012348
VERSION     V012348
KEYWORDS    CRISPRi library backbone
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8888)
  AUTHORS   Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead
            EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M,
            Weissman JS.
  TITLE     Genome-Scale CRISPR-Mediated Control of Gene Repression and
            Activation.
  JOURNAL   Cell 2014;159:647-61.
   PUBMED   25307932
REFERENCE   2  (bases 1 to 8888)
  AUTHORS   Weissman Lab / Addgene #62217
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8888)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell";
            date: "2014"; volume: "159"; pages: "647-61"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        247..626
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..829
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             844..1024
                     /label="LTR (truncated)"
                     /note="LTR (truncated)"
                     /note="truncated long terminal repeat (LTR) from HIV-1"
     misc_feature    1071..1196
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1695..1928
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2113..2157
                     /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             2306..2347
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2455..2572
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2631..2943
                     /note="U6 promoter"
                     /note="modified RNA polymerase III promoter for mouse U6
                     snRNA"
     protein_bind    2825..2858
                     /label="Cre recombinase binding site"
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     misc_feature    2965..3050
                     /label="gRNA scaffold"
                     /note="gRNA scaffold"
                     /note="modified guide RNA scaffold for the Streptococcus
                     pyogenes CRISPR/Cas9 system"
     terminator      3051..3056
                     /note="polIII terminator"
                     /note="RNA polymerase III transcription terminator"
     promoter        3114..4286
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             4298..4894
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     CDS             4904..4957
                     /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             4970..5665
                     /label="TagBFP"
                     /note="monomeric blue fluorescent protein"
     protein_bind    complement(5688..5721)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5777..6365
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(6368..6384)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             6894..7074
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7136..7724)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7898..8755)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8756..8860)
                     /label="AmpR promoter"