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Plasmid Files

pBiEx™-1

Vector for high-level expression in both bacterial and insect cells of proteins with a cleavable N-terminal 6xHis-S-Tag cassette plus a C-terminal HSV tag.

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pBiEx-1 Sequence and MappBiEx-1.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
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 BspQI - SapI (5226) PciI (5109) DrdI (5007) AhdI (4221) BsaI (4155) ScaI (3740) XmnI (3621) EcoNI (3232) PfoI (3196) BstAPI (3088) BstEII (2582) BspDI * - ClaI * (207) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BseRI (781) BsiWI (784) SacII (1085) T7 promoter RBS NcoI (1177) ATG 6xHis thrombin site BglII (1321) SrfI (1333) BamHI (1340) MfeI (1346) Eco53kI (1355) SacI (1357) BsrGI (1360) AscI (1368) PstI - SbfI (1378) SalI (1380) Acc65I (1386) AgeI (1389) KpnI (1390) BstBI (1395) HindIII (1398) NotI (1405) BstZ17I (1420) PmlI (1425) PaeR7I - XhoI (1467) DraIII (1478) BbvCI - Bpu10I (1488) BlpI (1525) EcoO109I (1552) Bsu36I (1602) PacI (1631) PflFI - Tth111I (1855) KasI (2125) NarI * (2126) SfoI (2127) PluTI (2129) BsmBI (2149) HpaI (2262) PspOMI (2557) ApaI (2561) pBiEx™-1 5475 bp
BspQI  (5226)
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  (5226)
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.
PciI  (5109)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (5007)
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.
AhdI  (4221)
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.
BsaI  (4155)
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.
ScaI  (3740)
1 site
A G T A C T T C A T G A
XmnI  (3621)
1 site
G A A N N N N T T C C T T N N N N A A G
EcoNI  (3232)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
PfoI  (3196)
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.
BstAPI  (3088)
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.
BstEII  (2582)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BspDI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PsiI  (409)
1 site
T T A T A A A A T A T T
NheI  (606)
1 site
G C T A G C C G A T C G
BmtI  (610)
1 site
G C T A G C C G A T C G
SphI  (614)
1 site
G C A T G C C G T A C G
BsmI  (775)
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.
BseRI  (781)
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.
BsiWI  (784)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
SacII  (1085)
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.
NcoI  (1177)
1 site
C C A T G G G G T A C C
BglII  (1321)
1 site
A G A T C T T C T A G A
SrfI  (1333)
1 site
G C C C G G G C C G G G C C C G
BamHI  (1340)
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.
MfeI  (1346)
1 site
C A A T T G G T T A A C
Eco53kI  (1355)
1 site
G A G C T C C T C G A G
SacI  (1357)
1 site
G A G C T C C T C G A G
BsrGI  (1360)
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.
AscI  (1368)
1 site
G G C G C G C C C C G C G C G G
PstI  (1378)
1 site
C T G C A G G A C G T C
SbfI  (1378)
1 site
C C T G C A G G G G A C G T C C
SalI  (1380)
1 site
G T C G A C C A G C T G
Acc65I  (1386)
1 site
G G T A C C C C A T G G
AgeI  (1389)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
KpnI  (1390)
1 site
G G T A C C C C A T G G
BstBI  (1395)
1 site
T T C G A A A A G C T T
HindIII  (1398)
1 site
A A G C T T T T C G A A
NotI  (1405)
1 site
G C G G C C G C C G C C G G C G
BstZ17I  (1420)
1 site
G T A T A C C A T A T G
PmlI  (1425)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
PaeR7I  (1467)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1467)
1 site
C T C G A G G A G C T C
DraIII  (1478)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BbvCI  (1488)
1 site
C C T C A G C G G A G T C G
Bpu10I  (1488)
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.
BlpI  (1525)
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.
EcoO109I  (1552)
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.
Bsu36I  (1602)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PacI  (1631)
1 site
T T A A T T A A A A T T A A T T
PflFI  (1855)
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  (1855)
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.
KasI  (2125)
1 site
G G C G C C C C G C G G
NarI  (2126)
1 site
G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (2127)
1 site
G G C G C C C C G C G G
PluTI  (2129)
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.
BsmBI  (2149)
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.
HpaI  (2262)
1 site
G T T A A C C A A T T G
PspOMI  (2557)
1 site
G G G C C C C C C G G G
ApaI  (2561)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
lacI
2037 .. 3119  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
lacI
2037 .. 3119  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
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).
AmpR
3434 .. 4294  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3434 .. 3502  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3434 .. 4294  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3503 .. 4294  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3434 .. 4294  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
IE1 promoter
487 .. 1078  =  592 bp
promoter of the ie1 gene from the baculovirus
Autographa californica
IE1 promoter
487 .. 1078  =  592 bp
promoter of the ie1 gene from the baculovirus
Autographa californica
ori
4465 .. 5053  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
4465 .. 5053  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hr5 enhancer
1 .. 483  =  483 bp
baculovirus early transcription enhancer
hr5 enhancer
1 .. 483  =  483 bp
baculovirus early transcription enhancer
MCS
1321 .. 1428  =  108 bp
multiple cloning site
MCS
1321 .. 1428  =  108 bp
multiple cloning site
AmpR promoter
3329 .. 3433  =  105 bp
AmpR promoter
3329 .. 3433  =  105 bp
lacI promoter
3120 .. 3197  =  78 bp
lacI promoter
3120 .. 3197  =  78 bp
T7 terminator
1536 .. 1583  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
1536 .. 1583  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
S-Tag
1212 .. 1256  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
S-Tag
1212 .. 1256  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
HSV tag
1434 .. 1466  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
1434 .. 1466  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
lac operator
1111 .. 1135  =  25 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
1111 .. 1135  =  25 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
1092 .. 1110  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1092 .. 1110  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
1185 .. 1202  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1185 .. 1202  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
thrombin site
1272 .. 1289  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
1272 .. 1289  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
RBS
1166 .. 1171  =  6 bp
ribosome binding site
RBS
1166 .. 1171  =  6 bp
ribosome binding site
ATG
1179 .. 1181  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1179 .. 1181  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
enterokinase site
1308 .. 1322  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
1308 .. 1322  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
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