pRANGER-BTB-1

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

Sequence Author: Lucigen

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HincII (0) EcoRI (3776) AflIII - MluI (3605) BstEII (3547) AgeI (3544) BspQI - EarI - SapI (3533) DrdI (3504) BspEI (3185) NruI (3178) SspI (3106) NspI (2838) AflII (2771) BsaBI (2711) AhdI (2418) BsaI (2352) AseI (2243) SfcI (2172) PvuI (2049) XcmI (2035) ScaI (1937) EcoRI (4) HindIII (10) SacII (103) PvuII (818) MscI (875) BbsI (1061) AfeI (1127) BclI * (1255) BseRI (1370) AlwNI - BstAPI (1450) PshAI (1511) SnaBI (1537) Bpu10I (1617) ApaLI (1746) XmnI (1818) pRANGER™-BTB-1 3784 bp
HincII  (0)
1 site
G T Y R A C C A R Y T G
EcoRI  (3776)
2 sites
G A A T T C C T T A A G
AflIII  (3605)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
MluI  (3605)
1 site
A C G C G T T G C G C A
BstEII  (3547)
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  (3544)
1 site
A C C G G T T G G C C A
BspQI  (3533)
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  (3533)
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  (3533)
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  (3504)
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.
BspEI  (3185)
1 site
T C C G G A A G G C C T
NruI  (3178)
1 site
T C G C G A A G C G C T
SspI  (3106)
1 site
A A T A T T T T A T A A
NspI  (2838)
1 site
R C A T G Y Y G T A C R
AflII  (2771)
1 site
C T T A A G G A A T T C
BsaBI  (2711)
1 site
G A T N N N N A T C C T A N N N N T A G
AhdI  (2418)
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  (2352)
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.
AseI  (2243)
1 site
A T T A A T T A A T T A
SfcI  (2172)
1 site
C T R Y A G G A Y R T C

Sticky ends from different SfcI sites may not be compatible.
SfcI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
PvuI  (2049)
1 site
C G A T C G G C T A G C
XcmI  (2035)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
ScaI  (1937)
1 site
A G T A C T T C A T G A
EcoRI  (4)
2 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.
PvuII  (818)
1 site
C A G C T G G T C G A C
MscI  (875)
1 site
T G G C C A A C C G G T
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.
ApaLI  (1746)
1 site
G T G C A C C A C G T G
XmnI  (1818)
1 site
G A A N N N N T T C C T T N N N N A A G
araC
2566 .. 3444  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
araC
2566 .. 3444  =  879 bp
292 amino acids  =  33.4 kDa
Product: L-arabinose regulatory protein
AmpR
1631 .. 2491  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   1631 .. 1699  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1631 .. 2491  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   1700 .. 2491  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1631 .. 2491  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
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
araBAD promoter
3590 .. 3755  =  166 bp
promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction
araBAD promoter
3590 .. 3755  =  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
2507 .. 2538  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
tonB terminator
2507 .. 2538  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
MCS
3776 .. 15  =  24 bp
multiple cloning site
MCS
3776 .. 15  =  24 bp
multiple cloning site
ORF:  3130 .. 3378  =  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 .. 2491  =  861 bp
ORF:  286 amino acids  =  31.5 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:  2095 .. 2397  =  303 bp
ORF:  100 amino acids  =  10.5 kDa
ORF:  2566 .. 3495  =  930 bp
ORF:  309 amino acids  =  35.2 kDa
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