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Plasmid Files

pTurboGFP-B

Vector for expressing TurboGFP in bacteria.

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pTurboGFP-B Sequence and MappTurboGFP-B.dna
Map and Sequence File   
Sequence Author:  Evrogen
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 BbsI (4089) AatII (4033) ZraI (4031) PvuI (3481) FspI (3333) AseI (3283) NmeAIII (3259) BglI (3231) BsaI (3172) AhdI (3111) AlwNI (2634) AflIII - PciI (2218) BstAPI (2042) PaeR7I - XhoI (1) PsiI (49) MfeI (59) lac operator ATG BamHI (127) KasI (277) NarI (278) SfoI (279) PluTI (281) NgoMIV (469) EagI - NaeI (471) FseI (473) AarI (581) PstI (707) PspOMI (714) ApaI - BanII (718) HindIII (828) NheI (948) BmtI (952) Bpu10I (973) PvuII (1100) BspEI (1196) PasI (1432) MscI (1467) XbaI (1805) PfoI (1862) PflFI - Tth111I (1965) AccI (1990) BstZ17I (1991) NdeI (2041) pTurboGFP-B 4103 bp
BbsI  (4089)
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  (4033)
1 site
G A C G T C C T G C A G
ZraI  (4031)
1 site
G A C G T C C T G C A G
PvuI  (3481)
1 site
C G A T C G G C T A G C
FspI  (3333)
1 site
T G C G C A A C G C G T
AseI  (3283)
1 site
A T T A A T T A A T T A
NmeAIII  (3259)
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  (3231)
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.
BsaI  (3172)
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.
AhdI  (3111)
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  (2634)
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  (2218)
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  (2218)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BstAPI  (2042)
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.
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
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.
KasI  (277)
1 site
G G C G C C C C G C G G
NarI  (278)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (279)
1 site
G G C G C C C C G C G G
PluTI  (281)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
NgoMIV  (469)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
EagI  (471)
1 site
C G G C C G G C C G G C
NaeI  (471)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
FseI  (473)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
AarI  (581)
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.
PstI  (707)
1 site
C T G C A G G A C G T C
PspOMI  (714)
1 site
G G G C C C C C C G G G
ApaI  (718)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BanII  (718)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
HindIII  (828)
1 site
A A G C T T T T C G A A
NheI  (948)
1 site
G C T A G C C G A T C G
BmtI  (952)
1 site
G C T A G C C G A T C G
Bpu10I  (973)
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  (1100)
1 site
C A G C T G G T C G A C
BspEI  (1196)
1 site
T C C G G A A G G C C T
PasI  (1432)
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  (1467)
1 site
T G G C C A A C C G G T
XbaI  (1805)
1 site
T C T A G A A G A T C T
PfoI  (1862)
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  (1965)
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  (1965)
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  (1990)
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  (1991)
1 site
G T A T A C C A T A T G
NdeI  (2041)
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.
AmpR
3038 .. 3898  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3038 .. 3829  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3038 .. 3898  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3830 .. 3898  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3038 .. 3898  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
TurboGFP
133 .. 828  =  696 bp
231 amino acids  =  25.6 kDa
Product: improved green fluorescent protein from
Pontellina plumata
mammalian codon-optimized
TurboGFP
133 .. 828  =  696 bp
231 amino acids  =  25.6 kDa
Product: improved green fluorescent protein from
Pontellina plumata
mammalian codon-optimized
CmR
987 .. 1646  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
987 .. 1646  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
2279 .. 2867  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2279 .. 2867  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
AmpR promoter
3899 .. 4003  =  105 bp
AmpR promoter
3899 .. 4003  =  105 bp
lambda t0 terminator
849 .. 943  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
849 .. 943  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
1711 .. 1797  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
1711 .. 1797  =  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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