pCas-Guide-EF1a-CD4

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HomePlasmidsCRISPR PlasmidspCas-Guide-EF1a-CD4
CF3 Primer (128 .. 149) SV40 NLS SV40 NLS BamHI (334) BsmBI (384) pCas-Guide-EF1a-CD4 11,006 bp
BamHI  (334)
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  (384)
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
128 .. 149  =  22 annealed bases
Tm  =  56°C
Cas9
4170 .. 8273  =  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
4170 .. 8273  =  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
8274 .. 8294  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
8274 .. 8294  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
8301 .. 8321  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
8301 .. 8321  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Myc
8343 .. 8372  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
8343 .. 8372  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
8391 .. 8414  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
8391 .. 8414  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
CD4 (truncated)
1679 .. 2950  =  1272 bp
424 amino acids  =  47.0 kDa
2 segments
   Segment 1:  signal peptide  
   1679 .. 1753  =  75 bp
   25 amino acids  =  2.7 kDa
Product: truncated human CD4 cell surface glycoprotein lacking the cytoplasmic tail
CD4 (truncated)
1679 .. 2950  =  1272 bp
424 amino acids  =  47.0 kDa
2 segments
   Segment 2:  
   1754 .. 2950  =  1197 bp
   399 amino acids  =  44.3 kDa
Product: truncated human CD4 cell surface glycoprotein lacking the cytoplasmic tail
CD4 (truncated)
1679 .. 2950  =  1272 bp
424 amino acids  =  47.0 kDa
2 segments
Product: truncated human CD4 cell surface glycoprotein lacking the cytoplasmic tail
EF-1α promoter
480 .. 1658  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
480 .. 1658  =  1179 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
9967 .. 10,827  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   9967 .. 10,758  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9967 .. 10,827  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   10,759 .. 10,827  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9967 .. 10,827  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
hGH poly(A) signal
8461 .. 9083  =  623 bp
human growth hormone polyadenylation signal
hGH poly(A) signal
8461 .. 9083  =  623 bp
human growth hormone polyadenylation signal
ori
9204 .. 9792  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9204 .. 9792  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
3560 .. 3863  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
3560 .. 3863  =  304 bp
human cytomegalovirus immediate early enhancer
U6 promoter
90 .. 330  =  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
90 .. 330  =  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.
CMV promoter
3864 .. 4067  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
3864 .. 4067  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
gRNA scaffold
384 .. 459  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
384 .. 459  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
T7 promoter
4093 .. 4111  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4093 .. 4111  =  19 bp
promoter for bacteriophage T7 RNA polymerase
polIII terminator
460 .. 465  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
460 .. 465  =  6 bp
RNA polymerase III transcription terminator
EF-1α intron A
711 .. 1649  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
711 .. 1649  =  939 bp
intron upstream of the start codon of human EF-1α
ORF:  1036 .. 1473  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  1115 .. 1348  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  1679 .. 2959  =  1281 bp
ORF:  426 amino acids  =  47.1 kDa
ORF:  9173 .. 9529  =  357 bp
ORF:  118 amino acids  =  13.2 kDa
ORF:  10,097 .. 10,363  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4023 .. 8420  =  4398 bp
ORF:  1465 amino acids  =  169.8 kDa
ORF:  1974 .. 2228  =  255 bp
ORF:  84 amino acids  =  8.9 kDa
ORF:  4428 .. 4670  =  243 bp
ORF:  80 amino acids  =  9.6 kDa
ORF:  6222 .. 6566  =  345 bp
ORF:  114 amino acids  =  13.5 kDa
ORF:  6720 .. 7124  =  405 bp
ORF:  134 amino acids  =  15.6 kDa
ORF:  7593 .. 8216  =  624 bp
ORF:  207 amino acids  =  24.3 kDa
ORF:  2282 .. 2605  =  324 bp
ORF:  107 amino acids  =  12.0 kDa
ORF:  8825 .. 9049  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  778 .. 1398  =  621 bp
ORF:  206 amino acids  =  23.0 kDa
ORF:  2638 .. 2925  =  288 bp
ORF:  95 amino acids  =  10.4 kDa
ORF:  9967 .. 10,827  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

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pNW3
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pCFD2-dU6-2gRNA
pDCC6
pRGE31
gRNA_AAVS1-T1
pCFD3-dU6-3gRNA
pDD162 (Peft-3_Cas9 + Empty sgRNA)
pRGEB31
gRNA_AAVS1-T2
pCFD4-U6-1_U6-3tandemgRNAs
pDR274
pRGEB32
gRNA_Cloning Vector
pCLIP-All-EFS-Ametrine
Peft-3_cas9-SV40_NLS_tbb-2 3'UTR
pROSA26-Puro-DNR
gRNA_GFP-T1
pCLIP-All-EFS-Blast
pET-28b-Cas9-His
pRPR1_gRNA_handle_RPR1t
gRNA_GFP-T2
pCLIP-All-EFS-Puro
pET-Cas9-6xHis
pSECC
HBT-pcoCas9
pCLIP-All-EFS-tRFP
pET-Cas9-NLS-6xHis
pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s)
hCas9
pCLIP-All-EFS-ZsGreen
pET-NLS-Cas9-6xHis
pSLQ1658-dCas9-EGFP
hCas9_D10A
pCLIP-All-hCMV-Blast
pET28a Cas9-Cys
pSpCas9(BB)-2A-GFP (PX458)
JDS246
pCLIP-All-hCMV-Puro
pFGC-pcoCas9
pSpCas9(BB)-2A-Puro (PX459) V2.0
lenti dCAS-VP64_Blast
pCLIP-All-hCMV-tRFP
pGL3-U6-sgRNA-PGK-puromycin
pSpCas9n(BB)-2A-GFP (PX461)
lenti MS2-P65-HSF1_Hygro
pCLIP-All-hCMV-ZsGreen
pgRNA-bacteria
pSpCas9n(BB)-2A-Puro (PX462) V2.0
lenti sgRNA(MS2)_zeo backbone
pCLIP-All-Nickase(D10A)-EFS-Blast
pgRNA-humanized
pSpCas9n(BB) (PX460)
lentiCas9-Blast
pCLIP-All-Nickase(D10A)-EFS-Puro
pGuide
pSpCas9 (PX165)
lentiCas9-EGFP
pCLIP-All-Nickase(D10A)-EFS-tRFP
pGuide-it-sgRNA1 Vector (Linear)
pSPgRNA
lentiCas9n(D10A)-Blast
pCLIP-All-Nickase(D10A)-EFS-ZsGreen
pGuide-it-tdTomato Vector (Linear)
pSQT834
lentiCRISPR v2
pCLIP-All-Nickase(D10A)-hCMV-Puro
pGuide-it-ZsGreen1 Vector (Linear)
pSQT1313
lentiGuide-Puro
pCLIP-All-Nickase(D10A)-hCMV-tRFP
pHAGE EF1-alpha dCas9-KRAB
pST1374-N-NLS-flag-linker-Cas9-D10A
LSL-Cas9-Rosa26TV
pCLIP-All-Nickase(D10A)-hCMV-ZsGreen
pHAGE EF1-alpha dCas9-VP64
pST1374-N-NLS-flag-linker-Cas9-H840A
M-SPcas
pCLIP-All-Nickase(D10A)-TRE3G-Blast
pHAGE TRE dCas9
pST1374-NLS-flag-linker-Cas9
M-ST1cas
pCLIP-All-Nickase(D10A)-TRE3G-Puro
pHAGE TRE dCas9-KRAB
pT3TS-nCas9n
MLM3613
pCLIP-All-Nickase(D10A)-TRE3G-tRFP
pHAGE TRE dCas9-VP64
pT7-Cas9
MLM3636
pCLIP-All-Nickase(D10A)-TRE3G-ZsGreen
pHAGE-TO-dCas9-3XGFP
pT7-Cas9-Nickase
MLM3639
pCLIP-All-TRE3G-Blast
pHAGE-TO-dCas9-3XmCherry
pT7-gRNA
MS2-P65-HSF1_GFP
pCLIP-All-TRE3G-Puro
pHAGE-TO-nls-st1dCas9-3nls-3XTagBFP2
pT7-Guide-IVT
MSP1101
pCLIP-All-TRE3G-tRFP
pHAGE-TO-nmdCas9-3XGFP
pTDH3-dCas9
MSP2133
pCLIP-All-TRE3G-ZsGreen
pHAGE-TO-nmdCas9-3XmCherry
pTDH3-dCas9-Mxi1
MSP2135
pCLIP-Cas9(D10a)-Nickase-EFS-Puro
pHR-SFFV-dCas9-BFP
pTOL-hCMV-TET3G-Hygro Transactivator
MSP469
pCLIP-Cas9(D10a)-Nickase-EFS-Blast
pHR-SFFV-dCas9-BFP-KRAB
pTPGI_dCas9_VP64
p3s-Cas9HC
pCLIP-Cas9(D10a)-Nickase-EFS-tRFP
pHR-SFFV-KRAB-dCas9-P2A-mCherry
pU6-BbsI-chiRNA
p414-TEF1p-Cas9-CYC1t
pCLIP-Cas9(D10a)-Nickase-EFS-ZsGreen
pHRdSV40-dCas9-10xGCN4_v4-P2A-BFP
PU6_klp-12_sgRNA
p415-GalL-Cas9-CYC1t
pCLIP-Cas9(D10a)-Nickase-hCMV-Blast
pHRdSV40-NLS-dCas9-24xGCN4_v4-NLS-P2A-BFP-dWPRE
pU6-(BbsI)_CBh-Cas9-T2A-BFP-P2A-Ad4E1B
p426-SNR52p-gRNA.CAN1.Y-SUP4t
pCLIP-Cas9(D10a)-Nickase-hCMV-Puro
pHRdSV40-scFv-GCN4-sfGFP-VP64-GB1-NLS
pU6-(BbsI)_CBh-Cas9-T2A-mcherry-P2A-Ad4E4orf6
pAAV-Guide-it-1 Vector (Linear)
pCLIP-Cas9(D10a)-Nickase-hCMV-tRFP
pHSN401
pUC119-gRNA
pAAV-Guide-it-Down Vector (Linear)
pCLIP-Cas9(D10a)-Nickase-hCMV-ZsGreen
pHsp70-Cas9
pUC57-sgRNA expression vector
pAAV-Guide-it-Up Vector
pCLIP-Cas9(D10a)-Nickase-TRE3G-Blast
pICH41308_hCas9
Puro-Cas9 donor
pAAVS1-BSD-DNR
pCLIP-Cas9(D10a)-Nickase-TRE3G-Puro
pICH47742_2x35S-5'UTR-hCas9(STOP)-NOST
pWJ66
pAAVS1-Puro-DNR
pCLIP-Cas9(D10a)-Nickase-TRE3G-tRFP
piCRg Entry
pWJ68
pAC2-dual-dCas9VP48-sgExpression
pCLIP-Cas9(D10a)-Nickase-TRE3G-ZsGreen
pICSL01009_AtU6p
pwtCas9-bacteria
pAc-sgRNA-Cas9
pCLIP-Cas9-Nuclease-EFS-Blast
pK7WGF2_hCas9
pX260
pAC149-pCR8-dCas9VP160
pCLIP-Cas9-Nuclease-EFS-Puro
pKLV-U6gRNA(BbsI)-PGKpuro2ABFP
pX330
pAC152-dual-dCas9VP64-sgExpression
pCLIP-Cas9-Nuclease-EFS-tRFP
pL-CRISPR.EFS.GFP
pX330A-1x2
pAC153-dual-dCas9VP96-sgExpression
pCLIP-Cas9-Nuclease-EFS-ZsGreen
pL-CRISPR.EFS.tRFP
pX330S-2
pAC154-dual-dCas9VP160-sgExpression
pCLIP-Cas9-Nuclease-hCMV-Blast
pL-CRISPR.SFFV.GFP
pX334
pAC155-pCR8-sgExpression
pCLIP-Cas9-Nuclease-hCMV-Puro
pL-CRISPR.SFFV.tRFP
pX335
pAC84-pCR8-dCas9
pCLIP-Cas9-Nuclease-hCMV-tRFP
pLenti-Cas9
PX551
pAC91-pmax-dCas9VP64
pCLIP-Cas9-Nuclease-hCMV-ZsGreen
pLenti-Cas9-IRES-Puro
PX552
pAC93-pmax-dCas9VP160
pCLIP-Cas9-Nuclease-TRE3G-Blast
pLenti-Cas9-P2A-tGFP
pX600
pAC94-pmax-dCas9VP160-2A-puro
pCLIP-Cas9-Nuclease-TRE3G-Puro
pLenti-Cas-Guide
pX601
pAC95-pmax-dCas9VP160-2A-neo
pCLIP-Cas9-Nuclease-TRE3G-tRFP
pLenti-EF1a-Cas9-IRES-Puro
pX602
pACT2-CAS9c
pCLIP-Cas9-Nuclease-TRE3G-ZsGreen
pLenti-Guide-Puro
PX851
pBS-Hsp70-Cas9
pCLIP-gRNA-EFS-Blast
pLKO.1-puro U6 sgRNA BfuAI stuffer
PX852
pCAG-T3-hCAS-pA
pCLIP-gRNA-EFS-Puro
pLSC-5
PX853
pCAG-T3-hCasD10A-pA
pCLIP-gRNA-EFS-tRFP
pLV hUbC-Cas9-T2A-GFP
PX854
pCas
pCLIP-gRNA-EFS-ZsGreen
pLV hUbC-dCas9 VP64-T2A-GFP
PX855
pCas-Guide-AAVS1
pCLIP-gRNA-hCMV-Blast
pLV hUbC-dCas9-T2A-GFP
PX856
pCas-Guide-CRISPRa
pCLIP-gRNA-hCMV-Puro
pLVX-hyg-sgRNA1 Vector (Linear)
pY010 (pcDNA3.1-hAsCpf1)
pCas-Guide-CRISPRi
pCLIP-gRNA-hCMV-tRFP
pLVX-puro-Cas9
pY016 (pcDNA3.1-hLbCpf1)
pCas-Guide-EF1a-CD4
pCLIP-gRNA-hCMV-ZsGreen
pLX-sgRNA
sgRNA(MS2) cloning backbone
pCas-Guide-EF1a-GFP
pCRCT
pMB66
SP-Cas9
pCas-Guide-Nickase
pCRISPomyces-1
pMB70
SP-dCas9-VPR
pCas-Guide-ROSA26
pCRISPomyces-2
pME-Cas9
VP12
pCas9
pCRISPR

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