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Plasmid Files

pSF-RecA-FLuc

Bacterial vector for expressing firefly luciferase from a strong constitutive recA promoter.

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pSF-RecA-FLuc Sequence and MappSF-RecA-FLuc.dna
Map and Sequence File   
Sequence Author:  Oxford Genetics
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 PmeI (5356) RsrII (5049) BsrDI (4766) PflFI - Tth111I (4651) PmeI (4308) PpuMI - SanDI (4242) AscI - BssHII (4151) FseI (4005) SwaI (3899) AlwNI (3426) ApaLI (3324) SwaI (3006) PacI (2880) BstEII (2854) 3' β- globin insulator AsiSI (5) BglII (232) XcmI (294) BsaI (306) recA ( Δ LexA) promoter BglII (428) EagI - NotI (470) HindIII (481) Eco53kI (491) SacI (493) EcoRI (497) RBS Acc65I (509) KpnI (513) NcoI (517) BbsI (577) PvuII (593) ScaI (684) BsrGI (1009) AleI (1017) FspI (1341) KasI (1466) NarI (1467) SfoI (1468) PluTI (1470) BlpI (1473) AgeI (1650) DraIII (1665) SexAI * (1858) BpmI (1899) XbaI (2167) BseRI - BsgI (2170) stop codons BspDI - ClaI (2207) BamHI (2216) StuI (2226) NheI (2232) BmtI (2236) stop codons AanI - PsiI (2382) BglI (2571) PstI - SbfI (2748) pSF-RecA-FLuc 5475 bp
PmeI  (5356)
2 sites
G T T T A A A C C A A A T T T G
RsrII  (5049)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BsrDI  (4766)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (4651)
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  (4651)
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.
PmeI  (4308)
2 sites
G T T T A A A C C A A A T T T G
PpuMI  (4242)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
SanDI  (4242)
1 site
G G G W C C C C C C W G G G

Sticky ends from different SanDI sites may not be compatible.
AscI  (4151)
1 site
G G C G C G C C C C G C G C G G
BssHII  (4151)
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.
FseI  (4005)
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.
SwaI  (3899)
2 sites
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
AlwNI  (3426)
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.
ApaLI  (3324)
1 site
G T G C A C C A C G T G
SwaI  (3006)
2 sites
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
PacI  (2880)
1 site
T T A A T T A A A A T T A A T T
BstEII  (2854)
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.
AsiSI  (5)
1 site
G C G A T C G C C G C T A G C G
BglII  (232)
2 sites
A G A T C T T C T A G A
XcmI  (294)
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.
BsaI  (306)
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.
BglII  (428)
2 sites
A G A T C T T C T A G A
EagI  (470)
1 site
C G G C C G G C C G G C
NotI  (470)
1 site
G C G G C C G C C G C C G G C G
HindIII  (481)
1 site
A A G C T T T T C G A A
Eco53kI  (491)
1 site
G A G C T C C T C G A G
SacI  (493)
1 site
G A G C T C C T C G A G
EcoRI  (497)
1 site
G A A T T C C T T A A G
Acc65I  (509)
1 site
G G T A C C C C A T G G
KpnI  (513)
1 site
G G T A C C C C A T G G
NcoI  (517)
1 site
C C A T G G G G T A C C
BbsI  (577)
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.
PvuII  (593)
1 site
C A G C T G G T C G A C
ScaI  (684)
1 site
A G T A C T T C A T G A
BsrGI  (1009)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
AleI  (1017)
1 site
C A C N N N N G T G G T G N N N N C A C
FspI  (1341)
1 site
T G C G C A A C G C G T
KasI  (1466)
1 site
G G C G C C C C G C G G
NarI  (1467)
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  (1468)
1 site
G G C G C C C C G C G G
PluTI  (1470)
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.
BlpI  (1473)
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.
AgeI  (1650)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
DraIII  (1665)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
SexAI  (1858)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
BpmI  (1899)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
XbaI  (2167)
1 site
T C T A G A A G A T C T
BseRI  (2170)
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.
BsgI  (2170)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BspDI  (2207)
1 site
A T C G A T T A G C T A
ClaI  (2207)
1 site
A T C G A T T A G C T A
BamHI  (2216)
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.
StuI  (2226)
1 site
A G G C C T T C C G G A
NheI  (2232)
1 site
G C T A G C C G A T C G
BmtI  (2236)
1 site
G C T A G C C G A T C G
AanI  (2382)
1 site
T T A T A A A A T A T T
PsiI  (2382)
1 site
T T A T A A A A T A T T
BglI  (2571)
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.
PstI  (2748)
1 site
C T G C A G G A C G T C
SbfI  (2748)
1 site
C C T G C A G G G G A C G T C C
luciferase
519 .. 2171  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luciferase gene
luciferase
519 .. 2171  =  1653 bp
550 amino acids  =  60.6 kDa
Product: firefly luciferase
synthetic luciferase gene
NeoR/KanR
4405 .. 5199  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
4405 .. 5199  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
3071 .. 3659  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3071 .. 3659  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
rrnG terminator
2593 .. 2729  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
2593 .. 2729  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
5212 .. 5348  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
rrnG terminator
5212 .. 5348  =  137 bp
transcription terminator from the E. coli ribosomal
RNA rrnG operon (Albrechtsen et al., 1991)
SV40 poly(A) signal
2281 .. 2402  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2281 .. 2402  =  122 bp
SV40 polyadenylation signal
5' β-globin insulator
18 .. 89  =  72 bp
insulator upstream of the human β-globin locus
(Farrell et al., 2002)
5' β-globin insulator
18 .. 89  =  72 bp
insulator upstream of the human β-globin locus
(Farrell et al., 2002)
3' β-globin insulator
2774 .. 2845  =  72 bp
insulator downstream of the human β-globin locus
(Farrell et al., 2002)
3' β-globin insulator
2774 .. 2845  =  72 bp
insulator downstream of the human β-globin locus
(Farrell et al., 2002)
recA (ΔLexA) promoter
359 .. 427  =  69 bp
constitutively active E. coli recA promoter lacking
the LexA binding site (Brent and Ptashne, 1981)
recA (ΔLexA) promoter
359 .. 427  =  69 bp
constitutively active E. coli recA promoter lacking
the LexA binding site (Brent and Ptashne, 1981)
T7 terminator
2517 .. 2563  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
2517 .. 2563  =  47 bp
transcription terminator for bacteriophage T7 RNA
polymerase
stop codons
2191 .. 2201  =  11 bp
stop codons in all three reading frames
stop codons
2191 .. 2201  =  11 bp
stop codons in all three reading frames
stop codons
2239 .. 2249  =  11 bp
stop codons in all three reading frames
stop codons
2239 .. 2249  =  11 bp
stop codons in all three reading frames
RBS
505 .. 510  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
505 .. 510  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
4392 .. 4397  =  6 bp
Shine-Dalgarno ribosome binding site
RBS
4392 .. 4397  =  6 bp
Shine-Dalgarno ribosome binding site
Kozak sequence
515 .. 521  =  7 bp
Kozak sequence
515 .. 521  =  7 bp
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