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Plasmid Files

pKillerRed-B

Vector for expressing KillerRed in bacteria.

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pKillerRed-B Sequence and MappKillerRed-B.dna
Map and Sequence File   
Sequence Author:  Evrogen
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 BbsI (4114) AatII (4058) ZraI (4056) XmnI (3735) PvuI (3506) FspI (3358) AseI (3308) NmeAIII (3284) BglI (3256) AhdI (3136) AlwNI (2659) PspFI (2551) BseYI (2547) BspQI - SapI (2127) BstAPI (2067) NdeI (2066) PaeR7I - XhoI (1) PsiI (49) MfeI (59) lac operator BseRI (115) ATG BsaBI * (126) BamHI (127) BanII (344) AarI - BfuAI - BspMI (490) PmlI (777) HindIII (853) BlpI (865) NheI (973) BmtI (977) BspEI (1221) PasI (1457) MscI (1492) XbaI (1830) PfoI (1887) PflFI - Tth111I (1990) AccI (2015) BstZ17I (2016) pKillerRed-B 4128 bp
BbsI  (4114)
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.
AatII  (4058)
1 site
G A C G T C C T G C A G
ZraI  (4056)
1 site
G A C G T C C T G C A G
XmnI  (3735)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (3506)
1 site
C G A T C G G C T A G C
FspI  (3358)
1 site
T G C G C A A C G C G T
AseI  (3308)
1 site
A T T A A T T A A T T A
NmeAIII  (3284)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BglI  (3256)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
AhdI  (3136)
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.
AlwNI  (2659)
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.
PspFI  (2551)
1 site
C C C A G C G G G T C G
BseYI  (2547)
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.
BspQI  (2127)
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  (2127)
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.
BstAPI  (2067)
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.
NdeI  (2066)
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.
PaeR7I  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1)
1 site
C T C G A G G A G C T C
PsiI  (49)
1 site
T T A T A A A A T A T T
MfeI  (59)
1 site
C A A T T G G T T A A C
BseRI  (115)
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.
BsaBI  (126)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BamHI  (127)
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.
BanII  (344)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
AarI  (490)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BfuAI  (490)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (490)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PmlI  (777)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
HindIII  (853)
1 site
A A G C T T T T C G A A
BlpI  (865)
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.
NheI  (973)
1 site
G C T A G C C G A T C G
BmtI  (977)
1 site
G C T A G C C G A T C G
BspEI  (1221)
1 site
T C C G G A A G G C C T
PasI  (1457)
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.
MscI  (1492)
1 site
T G G C C A A C C G G T
XbaI  (1830)
1 site
T C T A G A A G A T C T
PfoI  (1887)
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.
PflFI  (1990)
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  (1990)
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.
AccI  (2015)
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.
BstZ17I  (2016)
1 site
G T A T A C C A T A T G
AmpR
3063 .. 3923  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3063 .. 3854  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3063 .. 3923  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3855 .. 3923  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3063 .. 3923  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
KillerRed
133 .. 852  =  720 bp
239 amino acids  =  26.5 kDa
Product: red fluorescent protein KillerRed, a
genetically-encoded photosensitizer
mammalian codon-optimized
KillerRed
133 .. 852  =  720 bp
239 amino acids  =  26.5 kDa
Product: red fluorescent protein KillerRed, a
genetically-encoded photosensitizer
mammalian codon-optimized
CmR
1012 .. 1671  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1012 .. 1671  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2304 .. 2892  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2304 .. 2892  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
3924 .. 4028  =  105 bp
AmpR promoter
3924 .. 4028  =  105 bp
lambda t0 terminator
874 .. 968  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
874 .. 968  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1736 .. 1822  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
1736 .. 1822  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
T5 promoter
10 .. 54  =  45 bp
   Segment 1:  
   10 .. 24  =  15 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
   Segment 2:  -35  
   25 .. 30  =  6 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
   Segment 3:  
   31 .. 47  =  17 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
   Segment 4:  -10  
   48 .. 54  =  7 bp
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
T5 promoter
10 .. 54  =  45 bp
4 segments
bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator
lac operator
62 .. 78  =  17 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
62 .. 78  =  17 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).
ATG
115 .. 117  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
115 .. 117  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
lac operator
30 .. 46  =  17 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
30 .. 46  =  17 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).
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