pCRISPomyces-2

Streptomyces plasmid for expression of codon-optimized Cas9 and a single guide RNA (sgRNA).

Sequence Author: Zhao Lab / Addgene #61737

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HomePlasmidsCRISPR PlasmidspCRISPomyces-2
XbaI (5482) fd terminator BbsI (4889) lac operator BbsI (5350) lambda t0 terminator pCRISPomyces-2 10,995 bp
XbaI  (5482)
1 site		 T C T A G A A G A T C T
BbsI  (4889)
2 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  (5350)
2 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.
Cas9
303 .. 4409  =  4107 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
303 .. 4409  =  4107 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
pSG5 Rep
8055 .. 9497  =  1443 bp
480 amino acids  =  52.9 kDa
Product: replication initiator protein from the Streptomyces ghanaensis plasmid pSG5
mutated to remove the BbsI site
pSG5 Rep
8055 .. 9497  =  1443 bp
480 amino acids  =  52.9 kDa
Product: replication initiator protein from the Streptomyces ghanaensis plasmid pSG5
mutated to remove the BbsI site
ApmR
6316 .. 7116  =  801 bp
267 amino acids  =  28.8 kDa
Product: aminoglycoside 3-N-acetyltransferase type IV                
confers resistance to apramycin (Paget and Davies, 1996)
ApmR
6316 .. 7116  =  801 bp
267 amino acids  =  28.8 kDa
Product: aminoglycoside 3-N-acetyltransferase type IV                
confers resistance to apramycin (Paget and Davies, 1996)
ori
5524 .. 6112  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5524 .. 6112  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
rpsL promoter
1 .. 302  =  302 bp
promoter for the Xylanimonas cellulosilytica ribosomal protein S12 gene, mutated to remove the BbsI sites
rpsL promoter
1 .. 302  =  302 bp
promoter for the Xylanimonas cellulosilytica ribosomal protein S12 gene, mutated to remove the BbsI sites
lacZα
4902 .. 5198  =  297 bp
98 amino acids  =  11.0 kDa
Product: LacZα fragment of β-galactosidase
lacZα
4902 .. 5198  =  297 bp
98 amino acids  =  11.0 kDa
Product: LacZα fragment of β-galactosidase
gapdh promoter
4609 .. 4893  =  285 bp
glyceraldehyde-3-phosphate dehydrogenase promoter from Eggerthella lenta
gapdh promoter
4609 .. 4893  =  285 bp
glyceraldehyde-3-phosphate dehydrogenase promoter from Eggerthella lenta
oriT
10,665 .. 10,774  =  110 bp
incP origin of transfer
oriT
10,665 .. 10,774  =  110 bp
incP origin of transfer
gRNA scaffold
5351 .. 5426  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
gRNA scaffold
5351 .. 5426  =  76 bp
guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system
lambda t0 terminator
5432 .. 5482  =  51 bp
transcription terminator from phage lambda
lambda t0 terminator
5432 .. 5482  =  51 bp
transcription terminator from phage lambda
fd terminator
4571 .. 4608  =  38 bp
central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)
fd terminator
4571 .. 4608  =  38 bp
central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)
lac promoter
5242 .. 5272  =  31 bp
3 segments
   Segment 3:  -10  
   5242 .. 5248  =  7 bp
promoter for the E. coli lac operon
lac promoter
5242 .. 5272  =  31 bp
3 segments
   Segment 2:  
   5249 .. 5266  =  18 bp
promoter for the E. coli lac operon
lac promoter
5242 .. 5272  =  31 bp
3 segments
   Segment 1:  -35  
   5267 .. 5272  =  6 bp
promoter for the E. coli lac operon
lac promoter
5242 .. 5272  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
5218 .. 5234  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
5218 .. 5234  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
ORF:  6316 .. 7119  =  804 bp
ORF:  267 amino acids  =  28.8 kDa
ORF:  8035 .. 8259  =  225 bp
ORF:  74 amino acids  =  7.9 kDa
ORF:  9184 .. 9669  =  486 bp
ORF:  161 amino acids  =  17.9 kDa
ORF:  9826 .. 10,062  =  237 bp
ORF:  78 amino acids  =  8.4 kDa
ORF:  4772 .. 5026  =  255 bp
ORF:  84 amino acids  =  9.3 kDa
ORF:  303 .. 4409  =  4107 bp
ORF:  1368 amino acids  =  158.4 kDa
ORF:  4485 .. 4760  =  276 bp
ORF:  91 amino acids  =  9.9 kDa
ORF:  7380 .. 7892  =  513 bp
ORF:  170 amino acids  =  18.8 kDa
ORF:  7998 .. 8480  =  483 bp
ORF:  160 amino acids  =  17.1 kDa
ORF:  6532 .. 7125  =  594 bp
ORF:  197 amino acids  =  22.5 kDa
ORF:  4902 .. 5198  =  297 bp
ORF:  98 amino acids  =  11.0 kDa
ORF:  6378 .. 6686  =  309 bp
ORF:  102 amino acids  =  10.5 kDa
ORF:  7122 .. 8036  =  915 bp
ORF:  304 amino acids  =  33.5 kDa
ORF:  8055 .. 10,016  =  1962 bp
ORF:  653 amino acids  =  71.0 kDa
ORF:  17 .. 364  =  348 bp
ORF:  115 amino acids  =  12.2 kDa
ORF:  2216 .. 2665  =  450 bp
ORF:  149 amino acids  =  17.8 kDa
ORF:  8657 .. 8914  =  258 bp
ORF:  85 amino acids  =  9.1 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

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