Resources
Plasmid Files

pY010 (pcDNA3.1-hAsCpf1)

Zhang lab mammalian expression vector encoding a humanized version of the Acidaminococcus CRISPR-associated protein Cpf1.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pY010 (pcDNA3.1-hAsCpf1) Sequence and MappY010 (pcDNA3.1-hAsCpf1).dna
Map and Sequence File   
Sequence Author:  Zhang Lab / Addgene #69982
Download Free Trial Get SnapGene Viewer

 SgrDI (9462) SspI (9345) AhdI (8541) PciI (7651) BstBI (6982) RsrII (6816) BssHII (6697) PflFI - Tth111I (6418) SmaI (6113) TspMI - XmaI (6111) AvrII (6090) SexAI * (5857) DraIII (5567) BbsI (5253) XbaI (5027) PaeR7I - PspXI - XhoI (5021) NotI (5015) EcoRI (4988) 3xHA BamHI (4898) FseI (4878) MfeI (161) NruI (208) MluI (228) SpeI (249) SnaBI (590) Eco53kI (816) SacI (818) T7 promoter NheI (895) BmtI (899) AflII (908) HindIII (911) Acc65I (917) KpnI (921) SbfI (2190) BlpI (2219) AarI (2373) PmlI (3544) XcmI (4567) nucleoplasmin NLS pY010 (pcDNA3.1-hAsCpf1) 9464 bp
SgrDI  (9462)
1 site
C G T C G A C G G C A G C T G C
SspI  (9345)
1 site
A A T A T T T T A T A A
AhdI  (8541)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
PciI  (7651)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstBI  (6982)
1 site
T T C G A A A A G C T T
RsrII  (6816)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BssHII  (6697)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
PflFI  (6418)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (6418)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
SmaI  (6113)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (6111)
1 site
C C C G G G G G G C C C
XmaI  (6111)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
AvrII  (6090)
1 site
C C T A G G G G A T C C
SexAI  (5857)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
DraIII  (5567)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BbsI  (5253)
1 site
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.
XbaI  (5027)
1 site
T C T A G A A G A T C T
PaeR7I  (5021)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5021)
1 site
V C T C G A G B B G A G C T C V
XhoI  (5021)
1 site
C T C G A G G A G C T C
NotI  (5015)
1 site
G C G G C C G C C G C C G G C G
EcoRI  (4988)
1 site
G A A T T C C T T A A G
BamHI  (4898)
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.
FseI  (4878)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
MfeI  (161)
1 site
C A A T T G G T T A A C
NruI  (208)
1 site
T C G C G A A G C G C T
MluI  (228)
1 site
A C G C G T T G C G C A
SpeI  (249)
1 site
A C T A G T T G A T C A
SnaBI  (590)
1 site
T A C G T A A T G C A T
Eco53kI  (816)
1 site
G A G C T C C T C G A G
SacI  (818)
1 site
G A G C T C C T C G A G
NheI  (895)
1 site
G C T A G C C G A T C G
BmtI  (899)
1 site
G C T A G C C G A T C G
AflII  (908)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
HindIII  (911)
1 site
A A G C T T T T C G A A
Acc65I  (917)
1 site
G G T A C C C C A T G G
KpnI  (921)
1 site
G G T A C C C C A T G G
SbfI  (2190)
1 site
C C T G C A G G G G A C G T C C
BlpI  (2219)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
AarI  (2373)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
PmlI  (3544)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
XcmI  (4567)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
AsCpf1
929 .. 4849  =  3921 bp
1307 amino acids  =  151.2 kDa
Product: CRISPR-associated protein from
Acidaminococcus sp. BV3L6 (Zetsche et al., 2015)
AsCpf1
929 .. 4849  =  3921 bp
1307 amino acids  =  151.2 kDa
Product: CRISPR-associated protein from
Acidaminococcus sp. BV3L6 (Zetsche et al., 2015)
AmpR
8468 .. 9328  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   8468 .. 9259  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8468 .. 9328  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   9260 .. 9328  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8468 .. 9328  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
6172 .. 6966  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
6172 .. 6966  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
7712 .. 8297  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7712 .. 8297  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
5334 .. 5762  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
5334 .. 5762  =  429 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
235 .. 614  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
5776 .. 6105  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
5776 .. 6105  =  330 bp
SV40 enhancer and early promoter
bGH poly(A) signal
5064 .. 5288  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
5064 .. 5288  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
615 .. 818  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
7140 .. 7261  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
7140 .. 7261  =  122 bp
SV40 polyadenylation signal
AmpR promoter
9329 .. 9433  =  105 bp
AmpR promoter
9329 .. 9433  =  105 bp
3xHA
4904 .. 4984  =  81 bp
27 amino acids  =  3.3 kDa
Product: three tandem HA epitope tags
3xHA
4904 .. 4984  =  81 bp
27 amino acids  =  3.3 kDa
Product: three tandem HA epitope tags
nucleoplasmin NLS
4850 .. 4897  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
nucleoplasmin NLS
4850 .. 4897  =  48 bp
16 amino acids  =  1.7 kDa
Product: bipartite nuclear localization signal from
nucleoplasmin
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
863 .. 881  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
5956 .. 6091  =  136 bp
SV40 origin of replication
SV40 ori
5956 .. 6091  =  136 bp
SV40 origin of replication
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter