pGC Blue (linearized)

Linearized and dephosphorylated vector with 3'-C tails for GC cloning of PCR products and blue/white colony screening.

Sequence Author: Lucigen

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DraI - SwaI (1991) cat promoterBpu10I (1839) BbeI (1701) SfoI (1699) BsaHI - NarI (1698) KasI (1697) DrdI (1675) MscI (1618) PflFI - Tth111I (1581) BsmFI (1546) FaqI (1545) NmeAIII (1478) BsrDI (1469) BssHII (1299) BtgI - NcoI - StyI (1264) NaeI (1198) NgoMIV (1196) AvaII - RsrII (1181) TsoI (1173) BglII (1026) BspDI - ClaI (930) BspCNI (746) NheI (2003) BmtI (2007) lac operatorHindIII (2191) Acc65I (2197) KpnI (2201) Eco53kI (2205) SacI (2207) BamHI (2209) SpeI (2215) EcoRI (2240) <AhdI> (2251) <AhdI> (1) EcoRI (7) EcoRV (19) NotI (34) PaeR7I - PspXI - XhoI (40) XbaI (52) EcoO109I - PspOMI (58) ApaI (62) PvuI (213) HgaI (261) BpmI (287) AlwNI (619) ApaLI (714) BseMII (745) pGC™ Blue 2250 bp
DraI  (1991)
1 site
T T T A A A A A A T T T
SwaI  (1991)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
Bpu10I  (1839)
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.
BbeI  (1701)
1 site
G G C G C C C C G C G G

Cleavage may be enhanced when more than one copy of the BbeI recognition sequence is present.
SfoI  (1699)
1 site
G G C G C C C C G C G G
BsaHI  (1698)
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.
NarI  (1698)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (1697)
1 site
G G C G C C C C G C G G
DrdI  (1675)
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.
MscI  (1618)
1 site
T G G C C A A C C G G T
PflFI  (1581)
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  (1581)
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.
BsmFI  (1546)
1 site
G G G A C ( N ) 9-10 C C C T G ( N ) 9-10 ( N ) 4

Cleavage may be enhanced when more than one copy of the BsmFI recognition sequence is present.
Sticky ends from different BsmFI sites may not be compatible.
BsmFI is typically used at 65°C, but is 50% active at 37°C.
For full activity of FastDigest® BsmFI, add fresh S-adenosylmethionine (SAM).
BsmFI gradually loses activity when stored at -20°C.
FaqI  (1545)
1 site
G G G A C ( N ) 10 C C C T G ( N ) 10 ( N ) 4

Cleavage may be enhanced when more than one copy of the FaqI recognition sequence is present.
Sticky ends from different FaqI sites may not be compatible.
After cleavage, FaqI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
NmeAIII  (1478)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsrDI  (1469)
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.
BssHII  (1299)
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.
BtgI  (1264)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1264)
1 site
C C A T G G G G T A C C
StyI  (1264)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
NaeI  (1198)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (1196)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
AvaII  (1181)
1 site
G G W C C C C W G G

Sticky ends from different AvaII sites may not be compatible.
RsrII  (1181)
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.
TsoI  (1173)
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).
BglII  (1026)
1 site
A G A T C T T C T A G A
BspDI  (930)
1 site
A T C G A T T A G C T A
ClaI  (930)
1 site
A T C G A T T A G C T A
BspCNI  (746)
1 site
C T C A G ( N ) 7-8 N N G A G T C ( N ) 7-8

Sticky ends from different BspCNI sites may not be compatible.
BspCNI is typically used at 25°C, but is 75% active at 37°C.
For full activity, add fresh S-adenosylmethionine (SAM).
NheI  (2003)
1 site
G C T A G C C G A T C G
BmtI  (2007)
1 site
G C T A G C C G A T C G
HindIII  (2191)
1 site
A A G C T T T T C G A A
Acc65I  (2197)
1 site
G G T A C C C C A T G G
KpnI  (2201)
1 site
G G T A C C C C A T G G
Eco53kI  (2205)
1 site
G A G C T C C T C G A G
SacI  (2207)
1 site
G A G C T C C T C G A G
BamHI  (2209)
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.
SpeI  (2215)
1 site
A C T A G T T G A T C A
EcoRI  (2240)
2 sites
G A A T T C C T T A A G
End  (2251)
0 sites
Start  (1)
0 sites
EcoRI  (7)
2 sites
G A A T T C C T T A A G
EcoRV  (19)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
NotI  (34)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (40)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (40)
1 site
V C T C G A G B B G A G C T C V
XhoI  (40)
1 site
C T C G A G G A G C T C
XbaI  (52)
1 site
T C T A G