pBIND-ER(alpha)

Renilla luciferase vector encoding the Gal4 DNA-binding domain fused to an estrogen receptor ligand-binding domain.

Sequence Author: Promega

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MluI (7481) PciI (6504) AhdI (6334) BspDI - ClaI (5289) BstXI (5229) RsrII (5024) NaeI (5010) NgoMIV (5008) BssHII (4905) PluTI (4511) SfoI (4509) NarI (4508) KasI (4507) EcoO109I (4244) NruI (4060) Bsu36I (3985) EcoRV (3884) SpeI (152) CMV enhancer NdeI (387) T7 promoter NheI (1052) BmtI (1056) PaeR7I - XhoI (1300) GAL4 DNA binding domain AsiSI - PvuI (1558) BsmBI - Esp3I (2346) DraI - PmeI (2460) EcoRI (2465) TspMI - XmaI (2481) SmaI (2483) SbfI (2508) NotI (2603) PsiI (2726) MfeI (2755) CsiI - SexAI * (3100) SfiI (3286) AvrII (3333) DraIII (3587) pBIND-ER α 7605 bp
MluI  (7481)
1 site
A C G C G T T G C G C A
PciI  (6504)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (6334)
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  (5289)
1 site
A T C G A T T A G C T A
ClaI  (5289)
1 site
A T C G A T T A G C T A
BstXI  (5229)
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  (5024)
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  (5010)
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  (5008)
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  (4905)
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  (4511)
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  (4509)
1 site
G G C G C C C C G C G G
NarI  (4508)
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  (4507)
1 site
G G C G C C C C G C G G
EcoO109I  (4244)
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.
NruI  (4060)
1 site
T C G C G A A G C G C T
Bsu36I  (3985)
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.
EcoRV  (3884)
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.
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
BsmBI  (2346)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2346)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
DraI  (2460)
1 site
T T T A A A A A A T T T
PmeI  (2460)
1 site
G T T T A A A C C A A A T T T G
EcoRI  (2465)
1 site
G A A T T C C T T A A G
TspMI  (2481)
1 site
C C C G G G G G G C C C
XmaI  (2481)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2483)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SbfI  (2508)
1 site
C C T G C A G G G G A C G T C C
NotI  (2603)
1 site
G C G G C C G C C G C C G G C G
PsiI  (2726)
1 site
T T A T A A A A T A T T
MfeI  (2755)
1 site
C A A T T G G T T A A C
CsiI  (3100)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (3100)
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  (3286)
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.
AvrII  (3333)
1 site
C C T A G G G G A T C C
DraIII  (3587)
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.
hRluc
3399 .. 4331  =  933 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
hRluc
3399 .. 4331  =  933 bp
311 amino acids  =  36.0 kDa
Product: Renilla luciferase
human codon-optimized
NeoR/KanR
4383 .. 5174  =  792 bp
263 amino acids  =  28.9 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
4383 .. 5174  =  792 bp
263 amino acids  =  28.9 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
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
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
ER-LBD
1563 .. 2456  =  894 bp
297 amino acids  =  33.5 kDa
Product: estrogen nuclear receptor alpha ligand-binding domain
ER-LBD
1563 .. 2456  =  894 bp
297 amino acids  =  33.5 kDa
Product: estrogen nuclear receptor alpha ligand-binding domain
AmpR
5547 .. 6407  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5547 .. 5615  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5547 .. 6407  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5616 .. 6407  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5547 .. 6407