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pFN26A (BIND) hRluc-neo Flexi® Vector

Flexi® Vector encoding Renilla luciferase fused to a neomycin resistance gene. This vector is useful for testing transcriptional activation domains.

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pFN26A (BIND) hRluc-neo Flexi Vector Sequence and MappFN26A (BIND) hRluc-neo Flexi Vector.dna
Map and Sequence File   
Sequence Author:  Promega
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 BglII (7065) MluI (6946) PciI (5969) AhdI (5799) BspDI - ClaI (4754) poly(A) signal BstXI (4694) RsrII (4489) NaeI (4475) NgoMIV (4473) BssHII (4370) PluTI (3976) SfoI (3974) NarI (3973) KasI (3972) EcoO109I (3709) PasI (3638) SpeI (152) CMV enhancer NdeI (387) T7 promoter NheI (1052) BmtI (1056) PaeR7I - XhoI (1300) GAL4 DNA binding domain AsiSI - PvuI (1558) barnase DraI - PmeI (1925) EcoRI (1930) XbaI (1957) SbfI (1973) BlpI (2038) NotI (2068) PsiI (2191) MfeI (2220) SexAI * (2565) SV40 promoter SfiI (2751) StuI (2797) AvrII (2798) PflMI (2852) DraIII (3052) PfoI * (3144) EcoRV (3349) Bsu36I (3450) NruI (3525) pFN26A (BIND) hRluc-neo Flexi® Vector 7070 bp
BglII  (7065)
1 site
A G A T C T T C T A G A
MluI  (6946)
1 site
A C G C G T T G C G C A
PciI  (5969)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (5799)
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.
BspDI  (4754)
1 site
A T C G A T T A G C T A
ClaI  (4754)
1 site
A T C G A T T A G C T A
BstXI  (4694)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
RsrII  (4489)
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.
NaeI  (4475)
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.
NgoMIV  (4473)
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.
BssHII  (4370)
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.
PluTI  (3976)
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.
SfoI  (3974)
1 site
G G C G C C C C G C G G
NarI  (3973)
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.
KasI  (3972)
1 site
G G C G C C C C G C G G
EcoO109I  (3709)
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.
PasI  (3638)
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.
SpeI  (152)
1 site
A C T A G T T G A T C A
NdeI  (387)
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.
NheI  (1052)
1 site
G C T A G C C G A T C G
BmtI  (1056)
1 site
G C T A G C C G A T C G
PaeR7I  (1300)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1300)
1 site
C T C G A G G A G C T C
AsiSI  (1558)
1 site
G C G A T C G C C G C T A G C G
PvuI  (1558)
1 site
C G A T C G G C T A G C
DraI  (1925)
1 site
T T T A A A A A A T T T
PmeI  (1925)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (1930)
1 site
G A A T T C C T T A A G
XbaI  (1957)
1 site
T C T A G A A G A T C T
SbfI  (1973)
1 site
C C T G C A G G G G A C G T C C
BlpI  (2038)
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.
NotI  (2068)
1 site
G C G G C C G C C G C C G G C G
PsiI  (2191)
1 site
T T A T A A A A T A T T
MfeI  (2220)
1 site
C A A T T G G T T A A C
SexAI  (2565)
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.
SfiI  (2751)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
StuI  (2797)
1 site
A G G C C T T C C G G A
AvrII  (2798)
1 site
C C T A G G G G A T C C
PflMI  (2852)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
DraIII  (3052)
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.
PfoI  (3144)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
EcoRV  (3349)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
Bsu36I  (3450)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
NruI  (3525)
1 site
T C G C G A A G C G C T
hRluc
2864 .. 3796  =  933 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
hRluc
2864 .. 3796  =  933 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
AmpR
5012 .. 5872  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5012 .. 5080  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5012 .. 5872  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5081 .. 5872  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5012 .. 5872  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
3848 .. 4639  =  792 bp
263 amino acids  =  28.9 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
NeoR/KanR
3848 .. 4639  =  792 bp
263 amino acids  =  28.9 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin®)
ori
6030 .. 6618  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6030 .. 6618  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
GAL4 DNA binding domain
1083 .. 1520  =  438 bp
146 amino acids  =  16.8 kDa
Product: DNA binding domain of the GAL4
transcriptional activator
GAL4 DNA binding domain
1083 .. 1520  =  438 bp
146 amino acids  =  16.8 kDa
Product: DNA binding domain of the GAL4
transcriptional activator
SV40 promoter
2456 .. 2813  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2456 .. 2813  =  358 bp
SV40 enhancer and early promoter
barnase
1585 .. 1920  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus
amyloliquefaciens

The barnase gene is lethal in standard bacterial
transformation strains.
barnase
1585 .. 1920  =  336 bp
111 amino acids  =  12.5 kDa
Product: ribonuclease from Bacillus
amyloliquefaciens

The barnase gene is lethal in standard bacterial
transformation strains.
CMV enhancer
214 .. 517  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
214 .. 517  =  304 bp
human cytomegalovirus immediate early enhancer
cer region
6734 .. 7017  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
6734 .. 7017  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
518 .. 721  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
chimeric intron
857 .. 989  =  133 bp
chimera between introns from human β-globin and
immunoglobulin heavy chain genes
SV40 poly(A) signal
2090 .. 2211  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2090 .. 2211  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4907 .. 5011  =  105 bp
AmpR promoter
4907 .. 5011  =  105 bp
poly(A) signal
4703 .. 4751  =  49 bp
synthetic polyadenylation signal
poly(A) signal
4703 .. 4751  =  49 bp
synthetic polyadenylation signal
IgA linker
1527 .. 1553  =  27 bp
9 amino acids  =  880.0 Da
Product: peptide derived from a human Ig alpha
chain
IgA linker
1527 .. 1553  =  27 bp
9 amino acids  =  880.0 Da
Product: peptide derived from a human Ig alpha
chain
T7 promoter
1033 .. 1051  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1033 .. 1051  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
2664 .. 2799  =  136 bp
SV40 origin of replication
SV40 ori
2664 .. 2799  =  136 bp
SV40 origin of replication
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