下载GeneBank文件(.gb)

LOCUS       lentiCRISPR_v2.       14873 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Zhang lab lentiviral transfer plasmid, also known as pLentiCRISPR 
            v2, for CRISPR/Cas9 genome editing of mammalian cells with a single 
            guide RNA (sgRNA).
ACCESSION   .
VERSION     .
KEYWORDS    lentiCRISPR v2.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14873)
  AUTHORS   Sanjana NE, Shalem O, Zhang F.
  TITLE     Improved vectors and genome-wide libraries for CRISPR screening.
  JOURNAL   Nat. Methods 2014;11:783-4.
  PUBMED    25075903
REFERENCE   2  (bases 1 to 14873)
  AUTHORS   Zhang Lab / Addgene #52961
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 14873)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2014"; volume: "11"; pages: "783-4"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     Full sequence confirmed by Addgene.
FEATURES             Location/Qualifiers
     source          1..14873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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             2291..2332
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2608..2848
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    2853..4737
                     /label=filler
                     /note="filler"
                     /note="digest with BsmBI to replace the filler with an
                     sgRNA   "
     misc_RNA        4738..4813
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      4814..4819
                     /note="polIII terminator"
                     /note="RNA polymerase III transcription terminator"
     promoter        4875..5086
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5111..9214
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             9215..9262
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=bipartite nuclear localization signal from
                     nucl
                     /note="nucleoplasmin NLS"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9263..9286
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             9296..9352
                     /label=P2A
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             9353..9946
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     CDS             complement(10436..10447)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     LTR             10625..10858
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10890..11114
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11160..11588
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        11602..11931
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        11979..12026
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             12045..12416
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    12549..12682
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(12767..12797)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(12812..12833)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(13121..13709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(13883..14740)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(14741..14845)
                     /label=AmpR promoter