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Plasmid Files

pRANGER™-BTB-3

Broad host range expression vector with a chloramphenicol resistance marker and with the pBBR1 replicon for Gram-negative bacteria.

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pRANGER-BTB-3 Sequence and MappRANGER-BTB-3.dna
Map and Sequence File   
Sequence Author:  Lucigen
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 EcoRI (3578) BsaHI (3465) BstEII (3349) AgeI (3346) BspQI - EarI - SapI (3335) DrdI (3306) AcuI (3222) NruI (2980) NspI (2640) EciI (2614) AflII (2573) BsiHKAI (2529) BsaBI (2513) ScaI (2261) NcoI (2145) PasI (2076) BsmBI (2068) AclI (1931) EcoRI (1844) HincII (0) EcoRI (4) HindIII (10) SacII (103) FspI (826) BmrI (934) BbsI (1061) AfeI (1127) BclI * (1255) BseRI (1370) AlwNI - BstAPI (1450) PshAI (1511) SnaBI (1537) Bpu10I (1617) pRANGER™-BTB-3 3586 bp
EcoRI  (3578)
3 sites
G A A T T C C T T A A G
BsaHI  (3465)
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.
BstEII  (3349)
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.
AgeI  (3346)
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.
BspQI  (3335)
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.
EarI  (3335)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI
recognition sequence.
Sticky ends from different EarI sites may not be compatible.
SapI  (3335)
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.
DrdI  (3306)
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.
AcuI  (3222)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI
recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its
electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
NruI  (2980)
1 site
T C G C G A A G C G C T
NspI  (2640)
1 site
R C A T G Y Y G T A C R
EciI  (2614)
1 site
G G C G G A ( N ) 9 N N C C G C C T ( N ) 9

Sticky ends from different EciI sites may not be compatible.
AflII  (2573)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
BsiHKAI  (2529)
1 site
G W G C W C C W C G W G

Sticky ends from different BsiHKAI sites may not be compatible.
BsaBI  (2513)
1 site
G A T N N N N A T C C T A N N N N T A G
ScaI  (2261)
1 site
A G T A C T T C A T G A
NcoI  (2145)
1 site
C C A T G G G G T A C C
PasI  (2076)
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.
BsmBI  (2068)
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.
AclI  (1931)
1 site
A A C G T T T T G C A A
EcoRI  (1844)
3 sites
G A A T T C C T T A A G
HincII  (0)
1 site
G T Y R A C C A R Y T G
EcoRI  (4)
3 sites
G A A T T C C T T A A G
HindIII  (10)
1 site
A A G C T T T T C G A A
SacII  (103)
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.
FspI  (826)
1 site
T G C G C A A C G C G T
BmrI  (934)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
BbsI  (1061)
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.
AfeI  (1127)
1 site
A G C G C T T C G C G A
BclI  (1255)
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.
BseRI  (1370)
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.
AlwNI  (1450)
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.
BstAPI  (1450)
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.
PshAI  (1511)
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.
SnaBI  (1537)
1 site
T A C G T A A T G C A T
Bpu10I  (1617)
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.
araC
2368 .. 3246  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
araC
2368 .. 3246  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
pBBR1 Rep
872 .. 1534  =  663 bp
220 amino acids  =  24.4 kDa
Product: replication protein from the broad host
range plasmid pBBR1
pBBR1 Rep
872 .. 1534  =  663 bp
220 amino acids  =  24.4 kDa
Product: replication protein from the broad host
range plasmid pBBR1
CmR
1631 .. 2290  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1631 .. 2290  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
araBAD promoter
3392 .. 3557  =  166 bp
promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction
araBAD promoter
3392 .. 3557  =  166 bp
promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction
soxR terminator
60 .. 96  =  37 bp
bidirectional E. coli soxR transcription terminator
soxR terminator
60 .. 96  =  37 bp
bidirectional E. coli soxR transcription terminator
tonB terminator
2309 .. 2340  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
tonB terminator
2309 .. 2340  =  32 bp
bidirectional E. coli tonB-P14 transcription
terminator
MCS
3578 .. 15  =  24 bp
multiple cloning site
MCS
3578 .. 15  =  24 bp
multiple cloning site
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