lentiCas9-EGFP 载体 (V012330#)

载体名称:
lentiCas9-EGFP
载体抗性:
Ampicillin
载体长度:
13177 bp
载体类型:
CRISPR Plasmids
复制子:
ori
载体来源:
Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, Scott DA, Song
拷贝数:
High copy number
启动子:
EM7

lentiCas9-EGFP 载体载体图谱

lentiCas9-EGFP13177 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600CMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTSEF-1-alpha core promoterCas9nucleoplasmin NLSFLAGP2AEGFPWPRE5' LTR (truncated)bGH poly(A) signalf1 oriSV40 promoterEM7 promoterBleoRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoter

lentiCas9-EGFP 载体载体序列

LOCUS       lentiCas9-EGFP.       13177 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Zhang lab lentiviral vector for expressing human codon-optimized S. 
            pyogenes Cas9 plus enhanced GFP (EGFP).
ACCESSION   .
VERSION     .
KEYWORDS    lentiCas9-EGFP.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13177)
  AUTHORS   Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, Scott DA, Song 
            J, Pan JQ, Weissleder R, Lee H, Zhang F, Sharp PA.
  TITLE     Genome-wide CRISPR screen in a mouse model of tumor growth and 
            metastasis.
  JOURNAL   Cell 2015;160:1246-60.
  PUBMED    25748654
REFERENCE   2  (bases 1 to 13177)
  AUTHORS   
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 13177)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; 
            date: "2015"; volume: "160"; pages: "1246-60"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..13177
                     /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        2650..2861
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2886..6989
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             6990..7037
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=bipartite nuclear localization signal from
                     nucl
                     /note="nucleoplasmin NLS"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7038..7061
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             7071..7127
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label=2A peptide from porcine teschovirus-1
                     polyprotein
                     /note="P2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7128..7841
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    7869..8457
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             8982..9162
                     /label=5' LTR (truncated)
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    9194..9418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9464..9892
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        10017..10227
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        10283..10330
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             10349..10720
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    10853..10986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(11071..11101)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(11116..11137)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(11425..12013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(12187..13044)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(13045..13149)
                     /label=AmpR promoter

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