pRANGER-BTB-3

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

Sequence Author: Lucigen

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HincII (0) 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) 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
HincII  (0)
1 site
G T Y R A C C A R Y T G
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
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

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
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

Cleavage may be enhanced when more than one copy of the AcuI recognition sequence is present.
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
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.
BsmBI-v2 is an improved version of BsmBI.
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
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

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.
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
ORF:  2932 .. 3180  =  249 bp
ORF:  82 amino acids  =  9.3 kDa
ORF:  422 .. 766  =  345 bp
ORF:  114 amino acids  =  12.2 kDa
ORF:  860 .. 1534  =  675 bp
ORF:  224 amino acids  =  24.9 kDa
ORF:  1631 .. 2290  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  159 .. 398  =  240 bp
ORF:  79 amino acids  =  9.0 kDa
ORF:  1185 .. 1412  =  228 bp
ORF:  75 amino acids  =  8.3 kDa
ORF:  473 .. 712  =  240 bp
ORF:  79 amino acids  =  8.4 kDa
ORF:  776 .. 1291  =  516 bp
ORF:  171 amino acids  =  18.6 kDa
ORF:  1337 .. 1561  =  225 bp
ORF:  74 amino acids  =  8.3 kDa
ORF:  2368 .. 3297  =  930 bp
ORF:  309 amino acids  =  35.2 kDa
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