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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HomePlasmidsTA and GC Cloning VectorspGC Blue (linearized)
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 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
Cleavage may be enhanced when more than one copy of the HgaI recognition sequence is present.
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
3 segments
   Segment 1:  -35  
   2058 .. 2063  =  6 bp
promoter for the E. coli lac operon
lac promoter
2058 .. 2088  =  31 bp
3 segments
   Segment 2:  
   2064 .. 2081  =  18 bp
promoter for the E. coli lac operon
lac promoter
2058 .. 2088  =  31 bp
3 segments
   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
ORF:  1 .. 276  =  276 bp
ORF:  91 amino acids  =  10.5 kDa  (no start codon)
ORF:  1408 .. 1767  =  360 bp
ORF:  119 amino acids  =  12.5 kDa
ORF:  2 .. 241  =  240 bp
ORF:  79 amino acids  =  8.3 kDa  (no start codon)
ORF:  942 .. 1520  =  579 bp
ORF:  192 amino acids  =  21.5 kDa
ORF:  1271 .. 1657  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  1035 .. 1829  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
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Individual Sequences & Maps

Allele TA Vector (linearized)
pCR2.1-TOPO (linearized)
pJAZZ-OK GC (left arm)
pSpark TA
pBAD-TOPO (linearized)
pCR4-TOPO (linearized)
pJAZZ-OK GC (right arm)
pSpark TA Done
pBAD Thio-TOPO (linearized)
pCR8 GW TOPO (linearized)
pLenti6.3 V5-TOPO (linearized)
pSTBlue-1 AccepTor
pBlue-TOPO (linearized)
pCRII-TOPO (linearized)
pLenti7.3 V5-TOPO (linearized)
pT7Blue T-Vector
pcDNA3.1 CT-GFP-TOPO (linearized)
pDrive (linearized)
pLPS-T (linearized)
pTA Plus (linearized)
pcDNA3.1 NT-GFP-TOPO (linearized)
pEF6 V5-His-TOPO (linearized)
pLUG (linearized)
pTargeT (linearized)
pcDNA3.1 V5-His-TOPO (linearized)
pENTR5'-TOPO (linearized)
pLUG-Multi (linearized)
pTOP TA V2 (linearized)
pcDNA3.3-TOPO (linearized)
pET SUMO (linearized)
pMT V5-His-TOPO (linearized)
pTrcHis-TOPO (linearized)
pcDNA3.4-TOPO (linearized)
pETBlue-1 AccepTor
pOptiVEC-TOPO (linearized)
pTrcHis2-TOPO (linearized)
pcDNA4 HisMax-TOPO (linearized)
pEXP5-CT TOPO (linearized)
pPrime (linearized)
pTZ57R_T
pcDNA5 FRT TO-TOPO (linearized)
pEXP5-NT TOPO (linearized)
pQE-30 UA (linearized)
pUC57-T
pcDNA5 FRT V5-His-TOPO (linearized)
pGC Blue (linearized)
pSC-A-amp kan (linearized)
pYES2.1 V5-His-TOPO (linearized)
pcDNA6.2 C-EmGFP-GW TOPO (linearized)
pGEM-T (linearized)
pSecTag FRT V5-His-TOPO (linearized)
RBC T&A Cloning Vector (linearized)
pcDNA6.2 C-YFP-GW TOPO (linearized)
pGEM-T Easy (linearized)
pSMART GC HK (linearized)
T-Vector pMD19 (Simple)
pcDNA6.2 N-EmGFP-GW TOPO (linearized)
pGlow-TOPO (linearized)
pSMART GC LK (linearized)
T-Vector pMD20
pCR-XL-TOPO (linearized)
pIB V5-His-TOPO (linearized)

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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