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Plasmid Files

pCoHygro

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

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pCoHygro Sequence and MappCoHygro.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Sciences)
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 PfoI (46) SspI (4343) XmnI (4138) BsaI (3600) AhdI (3539) AlwNI (3062) AflIII - PciI (2646) BspQI - SapI (2530) BanII - SacI (2289) Eco53kI (2287) KpnI (2283) KasI (235) NarI (236) SfoI (237) PluTI (239) HindIII (399) SphI (409) SbfI (415) SalI (417) AccI (418) XbaI (423) PshAI (832) AsiSI (1169) RsrII (1213) SacII (1585) BbvCI (1870) BlpI (1956) PasI (2027) AleI (2056) AbsI - PaeR7I - PspXI - XhoI (2124) BsgI (2133) NheI (2134) BmtI (2138) BsmI (2223) Acc65I (2279) pCoHygro 4526 bp
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.
SspI  (4343)
1 site
A A T A T T T T A T A A
XmnI  (4138)
1 site
G A A N N N N T T C C T T N N N N A A G
BsaI  (3600)
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  (3539)
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  (3062)
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  (2646)
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  (2646)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (2530)
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  (2530)
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.
BanII  (2289)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (2289)
1 site
G A G C T C C T C G A G
Eco53kI  (2287)
1 site
G A G C T C C T C G A G
KpnI  (2283)
1 site
G G T A C C C C A T G G
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
SphI  (409)
1 site
G C A T G C C G T A C G
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.
XbaI  (423)
1 site
T C T A G A A G A T C T
PshAI  (832)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
AsiSI  (1169)
1 site
G C G A T C G C C G C T A G C G
RsrII  (1213)
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.
SacII  (1585)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
BbvCI  (1870)
1 site
C C T C A G C G G A G T C G
BlpI  (1956)
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  (2027)
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  (2056)
1 site
C A C N N N N G T G G T G N N N N C A C
AbsI  (2124)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (2124)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2124)
1 site
V C T C G A G B B G A G C T C V
XhoI  (2124)
1 site
C T C G A G G A G C T C
BsgI  (2133)
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  (2134)
1 site
G C T A G C C G A T C G
BmtI  (2138)
1 site
G C T A G C C G A T C G
BsmI  (2223)
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  (2279)
1 site
G G T A C C C C A T G G
HygR
805 .. 1830  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
HygR
805 .. 1830  =  1026 bp
341 amino acids  =  38.0 kDa
Product: hygromycin B phosphotransferase
confers resistance to hygromycin
AmpR
3466 .. 4326  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3466 .. 4257  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3466 .. 4326  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4258 .. 4326  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3466 .. 4326  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2707 .. 3295  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2707 .. 3295  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
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
2143 .. 2277  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2143 .. 2277  =  135 bp
SV40 polyadenylation signal
AmpR promoter
4327 .. 4431  =  105 bp
AmpR promoter
4327 .. 4431  =  105 bp
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