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Plasmid Files

pGC™ Blue (linearized)

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

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pGC Blue (linearized) Sequence and MappGC Blue (linearized).dna
Map and Sequence File   
Sequence Author:  Lucigen
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 DraI - SwaI (1991) cat promoter Bpu10I (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 operator HindIII (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

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.
BbeI  (1701)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the BbeI
recognition sequence.
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

Efficient cleavage requires at least two copies of the BsmFI
recognition sequence.
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).
...
FaqI  (1545)
1 site
G G G A C ( N ) 10 C C C T G ( N ) 10 ( N ) 4

Efficient cleavage requires at least two copies of the FaqI
recognition sequence.
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 A A G A T C T
EcoO109I  (58)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PspOMI  (58)
1 site
G G G C C C C C C G G G
ApaI  (62)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PvuI  (213)
1 site
C G A T C G G C T A G C
HgaI  (261)
1 site
G A C G C ( N ) 5 C T G C G ( N ) 5 ( N ) 5

Efficient cleavage requires at least two copies of the HgaI
recognition sequence.
Sticky ends from different HgaI sites may not be compatible.
BpmI  (287)
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.
AlwNI  (619)
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.
ApaLI  (714)
1 site
G T G C A C C A C G T G
BseMII  (745)
1 site
C T C A G ( N ) 8 N N G A G T C ( N ) 8

Sticky ends from different BseMII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
NeoR/KanR
1035 .. 1829  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
1035 .. 1829  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
ori
385 .. 972  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
385 .. 972  =  588 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
lacZα
1 .. 276  =  276 bp
91 amino acids  =  10.5 kDa
Product: LacZα fragment of β-galactosidase
lacZα
1 .. 276  =  276 bp
91 amino acids  =  10.5 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2132 .. 2251  =  120 bp
40 amino acids  =  4.3 kDa
Product: LacZα fragment of β-galactosidase
lacZα
2132 .. 2251  =  120 bp
40 amino acids  =  4.3 kDa
Product: LacZα fragment of β-galactosidase
cat promoter
1830 .. 1932  =  103 bp
promoter of the E. coli cat gene
cat promoter
1830 .. 1932  =  103 bp
promoter of the E. coli cat gene
tonB terminator
1933 .. 1964  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
tonB terminator
1933 .. 1964  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
lac promoter
2058 .. 2088  =  31 bp
   Segment 1:  -35  
   2058 .. 2063  =  6 bp
promoter for the E. coli lac operon
lac promoter
2058 .. 2088  =  31 bp
   Segment 2:  
   2064 .. 2081  =  18 bp
promoter for the E. coli lac operon
lac promoter
2058 .. 2088  =  31 bp
   Segment 3:  -10  
   2082 .. 2088  =  7 bp
promoter for the E. coli lac operon
lac promoter
2058 .. 2088  =  31 bp
3 segments
promoter for the E. coli lac operon
T3Te terminator
334 .. 363  =  30 bp
phage T3 early transcription terminator
T3Te terminator
334 .. 363  =  30 bp
phage T3 early transcription terminator
T7Te terminator
984 .. 1011  =  28 bp
phage T7 early transcription terminator
T7Te terminator
984 .. 1011  =  28 bp
phage T7 early transcription terminator
lac operator
2096 .. 2112  =  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
2096 .. 2112  =  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).
T7 promoter
71 .. 89  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
71 .. 89  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
2154 .. 2172  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
2154 .. 2172  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
M13 fwd
96 .. 112  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
96 .. 112  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2120 .. 2136  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2120 .. 2136  =  17 bp
common sequencing primer, one of multiple similar
variants
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