pCoBlast

Selection plasmid for insect cells, with a blasticidin resistance gene under control of the copia promoter.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsInsect Cell VectorspCoBlast
AatII (3842) ZraI (3840) SspI (3724) XmnI (3519) ScaI (3400) BpmI (2990) BsaI (2981) AhdI (2920) AlwNI (2443) AflIII - PciI (2027) PfoI (46) NdeI (184) BstAPI (185) KasI (235) NarI (236) SfoI (237) PluTI (239) HindIII (399) BfuAI - BspMI (404) SphI (409) PstI - SbfI (415) SalI (417) AccI (418) HincII (419) XbaI (423) BamHI (429) BbsI (882) EagI (913) PpuMI (956) NruI * (1028) BsaBI * (1095) BbvCI - Bpu10I (1251) BlpI (1337) PasI (1408) AleI (1437) AbsI - PaeR7I - PspXI - XhoI (1505) BsgI (1514) NheI (1515) BmtI (1519) BsmI (1604) Acc65I (1660) KpnI (1664) Eco53kI (1668) SacI (1670) EcoRI (1672) BspQI - SapI (1911) pCoBlast 3907 bp
AatII  (3842)
1 site		 G A C G T C C T G C A G
ZraI  (3840)
1 site		 G A C G T C C T G C A G
SspI  (3724)
1 site		 A A T A T T T T A T A A
XmnI  (3519)
1 site		 G A A N N N N T T C C T T N N N N A A G
ScaI  (3400)
1 site		 A G T A C T T C A T G A
BpmI  (2990)
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.
BsaI  (2981)
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.
AhdI  (2920)
1 site		 G A C N N N N N G T C C T G N N N N N C A G
The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AlwNI  (2443)
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.
AflIII  (2027)
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  (2027)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
PfoI  (46)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
NdeI  (184)
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.
BstAPI  (185)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
KasI  (235)
1 site		 G G C G C C C C G C G G
NarI  (236)
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.
SfoI  (237)
1 site		 G G C G C C C C G C G G
PluTI  (239)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
HindIII  (399)
1 site		 A A G C T T T T C G A A
BfuAI  (404)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (404)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SphI  (409)
1 site		 G C A T G C C G T A C G
PstI  (415)
1 site		 C T G C A G G A C G T C
SbfI  (415)
1 site		 C C T G C A G G G G A C G T C C
SalI  (417)
1 site		 G T C G A C C A G C T G
AccI  (418)
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.
HincII  (419)
1 site		 G T Y R A C C A R Y T G
XbaI  (423)
1 site		 T C T A G A A G A T C T
BamHI  (429)
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.
BbsI  (882)
1 site		 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.
EagI  (913)
1 site		 C G G C C G G C C G G C
PpuMI  (956)
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.
NruI  (1028)
1 site		 T C G C G A A G C G C T
* Blocked by Dam methylation.
BsaBI  (1095)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BbvCI  (1251)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (1251)
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.
BlpI  (1337)
1 site		 G C T N A G C C G A N T C G
Sticky ends from different BlpI sites may not be compatible.
PasI  (1408)
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.
AleI  (1437)
1 site		 C A C N N N N G T G G T G N N N N C A C
AbsI  (1505)
1 site		 C C T C G A G G G G A G C T C C
PaeR7I  (1505)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1505)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (1505)
1 site		 C T C G A G G A G C T C
BsgI  (1514)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
NheI  (1515)
1 site		 G C T A G C C G A T C G
BmtI  (1519)
1 site		 G C T A G C C G A T C G
BsmI  (1604)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
Acc65I  (1660)
1 site		 G G T A C C C C A T G G
KpnI  (1664)
1 site		 G G T A C C C C A T G G
Eco53kI  (1668)
1 site		 G A G C T C C T C G A G
SacI  (1670)
1 site		 G A G C T C C T C G A G
EcoRI  (1672)
1 site		 G A A T T C C T T A A G
BspQI  (1911)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different BspQI sites may not be compatible.
SapI  (1911)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
AmpR
2847 .. 3707  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2847 .. 3638  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2847 .. 3707  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   3639 .. 3707  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2847 .. 3707  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2088 .. 2676  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2088 .. 2676  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
BSD
797 .. 1195  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
797 .. 1195  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
copia promoter
501 .. 780  =  280 bp
strong promoter from the Drosophila transposable element copia (Sinclair et al., 1986)
copia promoter
501 .. 780  =  280 bp
strong promoter from the Drosophila transposable element copia (Sinclair et al., 1986)
SV40 poly(A) signal
1524 .. 1658  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1524 .. 1658  =  135 bp
SV40 polyadenylation signal
AmpR promoter
3708 .. 3812  =  105 bp
AmpR promoter
3708 .. 3812  =  105 bp
ORF:  451 .. 684  =  234 bp
ORF:  77 amino acids  =  9.0 kDa
ORF:  2977 .. 3243  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  797 .. 1195  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  1217 .. 1519  =  303 bp
ORF:  100 amino acids  =  10.5 kDa
ORF:  186 .. 479  =  294 bp
ORF:  97 amino acids  =  11.2 kDa
ORF:  785 .. 1099  =  315 bp
ORF:  104 amino acids  =  11.2 kDa
ORF:  2847 .. 3707  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

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