pLenti-Cas-Guide

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AmpR promoter FLAG SV40 NLS SV40 NLS cPPT/CTS CF3 Primer (2745 .. 2766) BamHI (2951) BsmBI (3001) polIII terminator CMV promoter T7 promoter pLenti-Cas-Guide 11,015 bp
BamHI  (2951)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BsmBI  (3001)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
CF3 Primer
22-mer  /  45% GC
1 binding site
2745 .. 2766  =  22 annealed bases
Tm  =  56°C
Cas9
3814 .. 7917  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
3814 .. 7917  =  4104 bp
1368 amino acids  =  158.4 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
SV40 NLS
7918 .. 7938  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
7918 .. 7938  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
7945 .. 7965  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
7945 .. 7965  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Myc
7987 .. 8016  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
7987 .. 8016  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
8035 .. 8058  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
8035 .. 8058  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
CmR
10,223 .. 10,882  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
10,223 .. 10,882  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
8199 .. 8787  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
8199 .. 8787  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
9558 .. 10,146  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9558 .. 10,146  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
3128 .. 3507  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
3128 .. 3507  =  380 bp
human cytomegalovirus immediate early enhancer
U6 promoter
2707 .. 2947  =  241 bp
Truncated RNA polymerase III promoter for human U6 snRNA used in pSIREN vectors such as pSIREN-RetroQ-ZsGreen1. Expected transcription start site is 8bp downstream of this feature.
U6 promoter
2707 .. 2947  =  241 bp
Truncated RNA polymerase III promoter for human U6 snRNA used in pSIREN vectors such as pSIREN-RetroQ-ZsGreen1. Expected transcription start site is 8bp downstream of this feature.
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
CMV promoter
3508 .. 3711  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
3508 .. 3711  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early promoter
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
9316 .. 9496  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
9316 .. 9496  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
10,883 .. 10,987  =  105 bp
AmpR promoter
10,883 .. 10,987  =  105 bp
gRNA scaffold
3001 .. 3076  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
3001 .. 3076  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
T7 promoter
3737 .. 3755  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3737 .. 3755  =  19 bp
promoter for bacteriophage T7 RNA polymerase
polIII terminator
3077 .. 3082  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
3077 .. 3082  =  6 bp
RNA polymerase III transcription terminator
ORF:  1561 .. 2448  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  3667 .. 8064  =  4398 bp
ORF:  1465 amino acids  =  169.8 kDa
ORF:  8302 .. 9342  =  1041 bp
ORF:  346 amino acids  =  38.3 kDa
ORF:  8289 .. 8843  =  555 bp
ORF:  184 amino acids  =  20.5 kDa
ORF:  1395 .. 1856  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  10,223 .. 10,882  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4072 .. 4314  =  243 bp
ORF:  80 amino acids  =  9.6 kDa
ORF:  5866 .. 6210  =  345 bp
ORF:  114 amino acids  =  13.5 kDa
ORF:  6364 .. 6768  =  405 bp
ORF:  134 amino acids  =  15.6 kDa
ORF:  7237 .. 7860  =  624 bp
ORF:  207 amino acids  =  24.3 kDa
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