pGGAselect

Cloning vector with two BsmBI restriction sites for Golden Gate assembly. See also pGGA.

Sequence Author: New England Biolabs

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AflIII - PciI (2100) DrdI (1998) BssSαI (1927) HaeII (1860) PspFI (1800) BseYI (1796) BsiHKAI (1790) ApaLI (1786) AlwNI (1691) AcuI (1558) BsaAI (1345) Bpu10I (1262) TsoI (1139) Forward (CW) Analysis Primer (234 .. 260) PstI - SbfI (238) AarI (241) PmeI (247) PspXI (284) BamHI (290) BstBI (300) BsaI (314) BsmBI - Esp3I (325) BbsI (336) BglII (356) BbsI (379) BsmBI - Esp3I (390) BsaI (401) AccI (411) PacI (423) BsaHI (428) ZraI (429) AatII (431) EagI - NotI (446) AseI (480) Reverse (CCW) Analysis Primer (457 .. 482) BanI (568) TatI (619) ScaI (621) SspI (726) MscI (771) PasI (803) PflMI * (809) BpmI (918) BspEI (1038) PvuII (1138) pGGAselect 2220 bp
AflIII  (2100)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2100)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (1998)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
BssSαI  (1927)
1 site
C A C G A G G T G C T C
HaeII  (1860)
1 site
R G C G C Y Y C G C G R
PspFI  (1800)
1 site
C C C A G C G G G T C G
BseYI  (1796)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
BsiHKAI  (1790)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
ApaLI  (1786)
1 site
G T G C A C C A C G T G
AlwNI  (1691)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AcuI  (1558)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
BsaAI  (1345)
1 site
Y A C G T R R T G C A Y
Bpu10I  (1262)
1 site
C C T N A G C G G A N T C G

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
TsoI  (1139)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
PstI  (238)
1 site
C T G C A G G A C G T C
SbfI  (238)
1 site
C C T G C A G G G G A C G T C C
AarI  (241)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its electrophoretic mobility.
PmeI  (247)
1 site
G T T T A A A C C A A A T T T G
PspXI  (284)
1 site
V C T C G A G B B G A G C T C V
BamHI  (290)
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.
BstBI  (300)
1 site
T T C G A A A A G C T T
BsaI  (314)
2 sites
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.
BsmBI  (325)
2 sites
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.
Esp3I  (325)
2 sites
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BbsI  (336)
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.
BglII  (356)
1 site
A G A T C T T C T A G A
BbsI  (379)
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.
BsmBI  (390)
2 sites
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.
Esp3I  (390)
2 sites
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BsaI  (401)
2 sites
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.
AccI  (411)
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.
PacI  (423)
1 site
T T A A T T A A A A T T A A T T
BsaHI  (428)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
ZraI  (429)
1 site
G A C G T C C T G C A G
AatII  (431)
1 site
G A C G T C C T G C A G
EagI  (446)
1 site
C G G C C G G C C G G C
NotI  (446)
1 site
G C G G C C G C C G C C G G C G
AseI  (480)
1 site
A T T A A T T A A T T A
BanI  (568)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
TatI  (619)
1 site
W G T A C W W C A T G W
ScaI  (621)
1 site
A G T A C T T C A T G A
SspI  (726)
1 site
A A T A T T T T A T A A
MscI  (771)
1 site
T G G C C A A C C G G T
PasI  (803)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (809)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BpmI  (918)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BspEI  (1038)
1 site
T C C G G A A G G C C T
PvuII  (1138)
1 site
C A G C T G G T C G A C
Forward (CW) Analysis Primer
27-mer  /  44% GC
1 binding site
234 .. 260  =  27 annealed bases
Tm  =  61°C
Reverse (CCW) Analysis Primer
26-mer  /  42% GC
1 binding site
457 .. 482  =  26 annealed bases
Tm  =  56°C
CmR
593 .. 1252  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
593 .. 1252  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
1456 .. 2044  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1456 .. 2044  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
cat promoter
1253 .. 1355  =  103 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
cat promoter
1253 .. 1355  =  103 bp
promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase
upstream MCS
233 .. 304  =  72 bp
upstream multiple cloning site
upstream MCS
233 .. 304  =  72 bp
upstream multiple cloning site
BsmBI insert
326 .. 394  =  69 bp
fragment released by digestion with BsmBI
BsmBI insert
326 .. 394  =  69 bp
fragment released by digestion with BsmBI
downstream MCS
410 .. 452  =  43 bp
downstream multiple cloning site
downstream MCS
410 .. 452  =  43 bp
downstream multiple cloning site
T7 promoter
464 .. 482  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
464 .. 482  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
254 .. 272  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
254 .. 272  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
ORF:  27 .. 263  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  593 .. 1252  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
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