pU6-(BbsI)_CBh-Cas9-T2A-BFP-P2A-Ad4E1B

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HomePlasmidsCRISPR PlasmidspU6-(BbsI)_CBh-Cas9-T2A-BFP-P2A-Ad4E1B
BbsI (251) BbsI (7076) BbsI (6835) BbsI (6813) BbsI (6415) BbsI (273) polIII terminator chicken β-actin promoter ATG SV40 NLS pU6-(BbsI)_CBh-Cas9-T2A-BFP-P2A-Ad4E1B 10,810 bp
BbsI  (251)
6 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BbsI  (7076)
6 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BbsI  (6835)
6 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BbsI  (6813)
6 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BbsI  (6415)
6 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BbsI  (273)
6 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
ATG
1257 .. 1259  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1257 .. 1259  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
3xFLAG
1260 .. 1325  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
3xFLAG
1260 .. 1325  =  66 bp
22 amino acids  =  2.7 kDa
Product: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site
SV40 NLS
1332 .. 1352  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
SV40 NLS
1332 .. 1352  =  21 bp
7 amino acids  =  883.1 Da
Product: nuclear localization signal of SV40 large T antigen
Cas9
1377 .. 5477  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
Cas9
1377 .. 5477  =  4101 bp
1367 amino acids  =  158.3 kDa
Product: Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system
generates RNA-guided double strand breaks in DNA
nucleoplasmin NLS
5478 .. 5525  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
nucleoplasmin NLS
5478 .. 5525  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from nucleoplasmin
T2A
5529 .. 5582  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
T2A
5529 .. 5582  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
EBFP2
5589 .. 6305  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 1:  
   5589 .. 5591  =  3 bp
   1 amino acid  =  149.2 Da
Product: enhanced blue variant of GFP (Ai et al., 2007)
mammalian codon-optimized
EBFP2
5589 .. 6305  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 2:  1a  
   5592 .. 5594  =  3 bp
   1 amino acid  =  117.1 Da
Product: enhanced blue variant of GFP (Ai et al., 2007)
mammalian codon-optimized
EBFP2
5589 .. 6305  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
   Segment 3:  
   5595 .. 6305  =  711 bp
   237 amino acids  =  26.7 kDa
Product: enhanced blue variant of GFP (Ai et al., 2007)
mammalian codon-optimized
EBFP2
5589 .. 6305  =  717 bp
239 amino acids  =  26.9 kDa
3 segments
Product: enhanced blue variant of GFP (Ai et al., 2007)
mammalian codon-optimized
P2A
6312 .. 6368  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1 polyprotein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
P2A
6312 .. 6368  =  57 bp
19 amino acids  =  2.0 kDa
Product: 2A peptide from porcine teschovirus-1 polyprotein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
E1B
6375 .. 7823  =  1449 bp
483 amino acids  =  54.2 kDa
Product: adenovirus 4 E1B 55K protein, which mediates the ubiquitination and proteasomal degradation of DNA ligase IV (Baker et al., 2007)
E1B
6375 .. 7823  =  1449 bp
483 amino acids  =  54.2 kDa
Product: adenovirus 4 E1B 55K protein, which mediates the ubiquitination and proteasomal degradation of DNA ligase IV (Baker et al., 2007)
AmpR
9130 .. 9990  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   9130 .. 9198  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9130 .. 9990  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   9199 .. 9990  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
9130 .. 9990  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
10,161 .. 10,749  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
10,161 .. 10,749  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
8288 .. 8743  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
8288 .. 8743  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
446 .. 731  =  286 bp
human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer
CMV enhancer
446 .. 731  =  286 bp
human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer
chicken β-actin promoter
733 .. 1010  =  278 bp
chicken β-actin promoter
733 .. 1010  =  278 bp
U6 promoter
7 .. 255  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
U6 promoter
7 .. 255  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
hybrid intron
1011 .. 1239  =  229 bp
hybrid between chicken β-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)
hybrid intron
1011 .. 1239  =  229 bp
hybrid between chicken β-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)
bGH poly(A) signal
7857 .. 8064  =  208 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
7857 .. 8064  =  208 bp
bovine growth hormone polyadenylation signal
AAV2 ITR
8073 .. 8213  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AAV2 ITR
8073 .. 8213  =  141 bp
inverted terminal repeat of adeno-associated virus serotype 2
AmpR promoter
9025 .. 9129  =  105 bp
AmpR promoter
9025 .. 9129  =  105 bp
gRNA scaffold
274 .. 349  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
274 .. 349  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
polIII terminator
350 .. 355  =  6 bp
RNA polymerase III transcription terminator
polIII terminator
350 .. 355  =  6 bp
RNA polymerase III transcription terminator
ORF:  1195 .. 1434  =  240 bp
ORF:  79 amino acids  =  9.1 kDa
ORF:  9130 .. 9990  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  641 .. 988  =  348 bp
ORF:  115 amino acids  =  12.1 kDa
ORF:  8087 .. 8311  =  225 bp
ORF:  74 amino acids  =  8.3 kDa
ORF:  831 .. 1142  =  312 bp
ORF:  103 amino acids  =  10.7 kDa
ORF:  1257 .. 7826  =  6570 bp
ORF:  2189 amino acids  =  250.3 kDa
ORF:  2723 .. 3016  =  294 bp
ORF:  97 amino acids  =  10.8 kDa
ORF:  3287 .. 3736  =  450 bp
ORF:  149 amino acids  =  17.3 kDa
ORF:  5462 .. 5731  =  270 bp
ORF:  89 amino acids  =  9.4 kDa
ORF:  7987 .. 8268  =  282 bp
ORF:  93 amino acids  =  10.4 kDa
ORF:  1236 .. 3893  =  2658 bp
ORF:  885 amino acids  =  93.1 kDa
ORF:  6570 .. 7115  =  546 bp
ORF:  181 amino acids  =  19.7 kDa
ORF:  7227 .. 7769  =  543 bp
ORF:  180 amino acids  =  19.7 kDa
ORF:  9594 .. 9860  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Individual Sequences & Maps

3xFLAG-dCas9 pCMV-7.1
pCas9_GFP
pCRISPRa-Enhancer
pME-Cas9-T2A-GFP
3xFLAG-dCas9 pMXs-neo
pCas9D10A_GFP
pCS2+hSpCas9
pMJ806
3xFLAG-dCas9 pMXs-puro
pCas-Guide
pCS2-Cas9
pMJ841
Cas9m4
pcDNA-dCas9
pCS2-nCas9n
pMJ920
Cas9m4-VP64
pcDNA-dCas9-p300 Core
pCW-Cas9
pMLM3705
CRISPRi library backbone
pcDNA-dCas9-p300 Core (D1399Y)
pdCas9
pMSCV-LTR-dCas9-p65AD-BFP
dCAS9-VP64_GFP
pcDNA-dCas9-VP64
pdCas9-bacteria
pMSCV-LTR-dCas9-VP64-BFP
eSpCas9(1.1)
pcDNA3.1-CibN-dCas9-CibN
pdCas9-humanized
pMZ374
GeneArt CRISPR Nuclease Vector - CD4 (linearized)
pCFD1-dU6-1gRNA
pdCas9 BFP-humanized
pNW3
GeneArt CRISPR Nuclease Vector - OFP (linearized)
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

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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