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Plasmid Files

pAcGFP1-C2

Vector for fusing AcGFP1 to the N-terminus of a partner protein. For other reading frames, use pAcGFP1‑C1 or pAcGFP1‑C3.

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pAcGFP1-C2 Sequence and MappAcGFP1-C2.dna
Map and Sequence File   
Sequence Author:  Clontech
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 PciI (4677) ApaLI (4363) BsaI (3748) RsrII (3275) BsrDI (2992) PflFI - Tth111I (2877) FspI (2861) BspDI * - ClaI * (2599) StuI (2580) BseRI (2577) SfiI (2534) SV40 promoter AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) AgeI (600) BstEII (795) Bpu10I (805) BssHII (939) EcoNI (1237) PpuMI (1242) PflMI (1311) BglII (1343) PaeR7I - XhoI (1347) Eco53kI (1352) SacI (1354) HindIII (1356) EcoRI (1363) PstI (1372) SalI (1373) AccI (1374) Acc65I (1379) KpnI (1383) SacII (1386) PspOMI (1387) TspMI - XmaI (1390) ApaI (1391) SmaI (1392) BamHI (1394) XbaI * (1406) stop codons BclI * (1416) MfeI (1509) HpaI (1522) MluI (1645) SexAI * (2348) pAcGFP1-C2 4735 bp
PciI  (4677)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (4363)
1 site
G T G C A C C A C G T G
BsaI  (3748)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
RsrII  (3275)
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.
BsrDI  (2992)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (2877)
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  (2877)
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.
FspI  (2861)
1 site
T G C G C A A C G C G T
BspDI  (2599)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2599)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2580)
1 site
A G G C C T T C C G G A
BseRI  (2577)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
SfiI  (2534)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
SnaBI  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
AgeI  (600)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
BstEII  (795)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
Bpu10I  (805)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BssHII  (939)
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.
EcoNI  (1237)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
PpuMI  (1242)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
PflMI  (1311)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BglII  (1343)
1 site
A G A T C T T C T A G A
PaeR7I  (1347)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1347)
1 site
C T C G A G G A G C T C
Eco53kI  (1352)
1 site
G A G C T C C T C G A G
SacI  (1354)
1 site
G A G C T C C T C G A G
HindIII  (1356)
1 site
A A G C T T T T C G A A
EcoRI  (1363)
1 site
G A A T T C C T T A A G
PstI  (1372)
1 site
C T G C A G G A C G T C
SalI  (1373)
1 site
G T C G A C C A G C T G
AccI  (1374)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI sites may not be compatible.
Acc65I  (1379)
1 site
G G T A C C C C A T G G
KpnI  (1383)
1 site
G G T A C C C C A T G G
SacII  (1386)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
PspOMI  (1387)
1 site
G G G C C C C C C G G G
TspMI  (1390)
1 site
C C C G G G G G G C C C
XmaI  (1390)
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.
ApaI  (1391)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (1392)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (1394)
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.
XbaI  (1406)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1416)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
MfeI  (1509)
1 site
C A A T T G G T T A A C
HpaI  (1522)
1 site
G T T A A C C A A T T G
MluI  (1645)
1 site
A C G C G T T G C G C A
SexAI  (2348)
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.
NeoR/KanR
2631 .. 3425  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
2631 .. 3425  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
AcGFP1
613 .. 1329  =  717 bp
239 amino acids  =  26.9 kDa
Product: Aequorea coerulescens GFP
mammalian codon-optimized
AcGFP1
613 .. 1329  =  717 bp
239 amino acids  =  26.9 kDa
Product: Aequorea coerulescens GFP
mammalian codon-optimized
ori
4033 .. 4621  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4033 .. 4621  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
1651 .. 2106  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
1651 .. 2106  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
SV40 promoter
2239 .. 2596  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2239 .. 2596  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
SV40 poly(A) signal
1523 .. 1644  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1523 .. 1644  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2133 .. 2237  =  105 bp
AmpR promoter
2133 .. 2237  =  105 bp
MCS
1343 .. 1399  =  57 bp
multiple cloning site of fluorescent protein plasmids
MCS
1343 .. 1399  =  57 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3657 .. 3704  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3657 .. 3704  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
stop codons
1408 .. 1418  =  11 bp
stop codons in all three reading frames
stop codons
1408 .. 1418  =  11 bp
stop codons in all three reading frames
SV40 ori
2447 .. 2582  =  136 bp
SV40 origin of replication
SV40 ori
2447 .. 2582  =  136 bp
SV40 origin of replication
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