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Plasmid Files

pMCSG79 (linearized)

Linearized transposon-containing vector for ligation-independent cloning (LIC), with a 6xHis-TEV leader, for use with the Bac-to-Bac® baculovirus system.

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pMCSG79 (linearized) Sequence and MappMCSG79 (linearized).dna
Map and Sequence File   
Sequence Author:  Midwest Center for Structural Genomics
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 NgoMIV (826) AvrII (339) BclI * (324) HpaI (188) MfeI (175) HindIII (60) SphI (52) PaeR7I - XhoI (42) PstI (39) XbaI (27) Start (0) End (4802) ATG NdeI (4747) BamHI (4731) BstZ17I (4583) AccI (4582) SnaBI (4580) BbsI (4456) BsrGI (4109) PflFI - Tth111I (3927) BsmBI (3881) BseRI (3660) EcoRV (3524) SacII (3467) BstXI (3416) MscI (3408) NaeI (828) BanII (860) BsaHI (1535) PvuI (1706) FspI (1852) BpmI (2006) AlwNI (2554) BseYI (2659) PspFI (2663) BspQI - SapI (3080) pMCSG79 4802 bp
NgoMIV  (826)
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.
AvrII  (339)
1 site
C C T A G G G G A T C C
BclI  (324)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
HpaI  (188)
1 site
G T T A A C C A A T T G
MfeI  (175)
1 site
C A A T T G G T T A A C
HindIII  (60)
1 site
A A G C T T T T C G A A
SphI  (52)
1 site
G C A T G C C G T A C G
PaeR7I  (42)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (42)
1 site
C T C G A G G A G C T C
PstI  (39)
1 site
C T G C A G G A C G T C
XbaI  (27)
1 site
T C T A G A A G A T C T
Start  (0)
0 sites
End  (4802)
0 sites
NdeI  (4747)
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.
BamHI  (4731)
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.
BstZ17I  (4583)
1 site
G T A T A C C A T A T G
AccI  (4582)
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.
SnaBI  (4580)
1 site
T A C G T A A T G C A T
BbsI  (4456)
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.
BsrGI  (4109)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
PflFI  (3927)
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  (3927)
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.
BsmBI  (3881)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BseRI  (3660)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
EcoRV  (3524)
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.
SacII  (3467)
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.
BstXI  (3416)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
MscI  (3408)
1 site
T G G C C A A C C G G T
NaeI  (828)
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.
BanII  (860)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BsaHI  (1535)
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.
PvuI  (1706)
1 site
C G A T C G G C T A G C
FspI  (1852)
1 site
T G C G C A A C G C G T
BpmI  (2006)
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  (2554)
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.
BseYI  (2659)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
PspFI  (2663)
1 site
C C C A G C G G G T C G
BspQI  (3080)
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  (3080)
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
1288 .. 2148  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1288 .. 1356  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1288 .. 2148  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1357 .. 2148  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1288 .. 2148  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
2319 .. 2907  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2319 .. 2907  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
GmR
3501 .. 4034  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
3501 .. 4034  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
701 .. 1156  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
701 .. 1156  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
Tn7R
3210 .. 3434  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
3210 .. 3434  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
352 .. 517  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
352 .. 517  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
189 .. 323  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
189 .. 323  =  135 bp
SV40 polyadenylation signal
AmpR promoter
1183 .. 1287  =  105 bp
AmpR promoter
1183 .. 1287  =  105 bp
polyhedrin promoter
4603 .. 4694  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
4603 .. 4694  =  92 bp
promoter for the baculovirus polyhedrin gene
Pc promoter
4223 .. 4251  =  29 bp
   Segment 3:  -10  
   4223 .. 4228  =  6 bp
class 1 integron promoter
Pc promoter
4223 .. 4251  =  29 bp
   Segment 2:  
   4229 .. 4245  =  17 bp
class 1 integron promoter
Pc promoter
4223 .. 4251  =  29 bp
   Segment 1:  -35  
   4246 .. 4251  =  6 bp
class 1 integron promoter
Pc promoter
4223 .. 4251  =  29 bp
3 segments
class 1 integron promoter
TEV site
4794 .. 4814  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
TEV site
4794 .. 4814  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease
recognition and cleavage site
6xHis
4752 .. 4769  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4752 .. 4769  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ATG
4749 .. 4751  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4749 .. 4751  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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