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Plasmid Files

pTurboYFP-B

Vector for expressing TurboYFP in bacteria.

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pTurboYFP-B.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Evrogen
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BbsI (4126) EcoO109I (4124) AatII (4070) ZraI (4068) PvuI (3518) FspI (3370) AseI (3320) BglI (3268) AhdI (3148) AlwNI (2671) AflIII - PciI (2255) BstAPI (2079) NdeI (2078) PaeR7I - XhoI (1) PsiI (49) MfeI (59) BseRI (115) BsaBI * (126) BamHI (127) SacII (222) SgrAI (280) AleI (310) SphI (390) BssHII (804) HindIII (865) NheI (985) BmtI (989) Bpu10I (1010) PvuII (1137) PasI (1469) NcoI - StyI (1538) XbaI (1842) PfoI (1899) PflFI - Tth111I (2002) AccI (2027) BstZ17I (2028) pTurboYFP-B 4140 bp
BbsI  (4126)
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.
EcoO109I  (4124)
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.
AatII  (4070)
1 site
G A C G T C C T G C A G
ZraI  (4068)
1 site
G A C G T C C T G C A G
PvuI  (3518)
1 site
C G A T C G G C T A G C
FspI  (3370)
1 site
T G C G C A A C G C G T
AseI  (3320)
1 site
A T T A A T T A A T T A
BglI  (3268)
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  (3148)
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  (2671)
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.
AflIII  (2255)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2255)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstAPI  (2079)
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  (2078)
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.
SacII  (222)
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.
SgrAI  (280)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
AleI  (310)
1 site
C A C N N N N G T G G T G N N N N C A C
SphI  (390)
1 site
G C A T G C C G T A C G
BssHII  (804)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
HindIII  (865)
1 site
A A G C T T T T C G A A
NheI  (985)
1 site
G C T A G C C G A T C G
BmtI  (989)
1 site
G C T A G C C G A T C G
Bpu10I  (1010)
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.
PvuII  (1137)
1 site
C A G C T G G T C G A C
PasI  (1469)
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.
NcoI  (1538)
1 site
C C A T G G G G T A C C
StyI  (1538)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
XbaI  (1842)
1 site
T C T A G A A G A T C T
PfoI  (1899)
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  (2002)
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  (2002)
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  (2027)
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  (2028)
1 site
G T A T A C C A T A T G
AmpR
3075 .. 3935  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3075 .. 3866  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3075 .. 3935  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3867 .. 3935  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3075 .. 3935  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
TurboYFP
133 .. 864  =  732 bp
243 amino acids  =  26.9 kDa
Product: enhanced derivative of PhiYFP
mammalian codon-optimized
TurboYFP
133 .. 864  =  732 bp
243 amino acids  =  26.9 kDa
Product: enhanced derivative of PhiYFP
mammalian codon-optimized
CmR
1024 .. 1683  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
1024 .. 1683  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2316 .. 2904  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2316 .. 2904  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
3936 .. 4040  =  105 bp
AmpR promoter
3936 .. 4040  =  105 bp
lambda t0 terminator
886 .. 980  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
886 .. 980  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1748 .. 1834  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
1748 .. 1834  =  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).
ORF:  115 .. 864  =  750 bp
ORF:  249 amino acids  =  27.4 kDa
ORF:  1024 .. 1683  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  3205 .. 3471  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3075 .. 3935  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
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