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Plasmid Files

pBIND-ERα

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

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pBIND-ER(alpha) Sequence and MappBIND-ER(alpha).dna
Map and Sequence File   
Sequence Author:  Promega
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 MluI (7481) PciI (6504) AhdI (6334) BspDI - ClaI (5289) poly(A) signal 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 (2346) DraI - PmeI (2460) EcoRI (2465) TspMI - XmaI (2481) SmaI (2483) SbfI (2508) NotI (2603) PsiI (2726) MfeI (2755) 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.
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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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
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
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
   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
   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  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
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®)
ori
6565 .. 7153  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6565 .. 7153  =  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
2991 .. 3348  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2991 .. 3348  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
214 .. 517  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
214 .. 517  =  304 bp
human cytomegalovirus immediate early enhancer
cer region
7269 .. 7552  =  284 bp
ColE1-derived recombination site that helps to
maintain plasmids as monomers
cer region
7269 .. 7552  =  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
2625 .. 2746  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2625 .. 2746  =  122 bp
SV40 polyadenylation signal
AmpR promoter
5442 .. 5546  =  105 bp
AmpR promoter
5442 .. 5546  =  105 bp
poly(A) signal
5238 .. 5286  =  49 bp
synthetic polyadenylation signal
poly(A) signal
5238 .. 5286  =  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
3199 .. 3334  =  136 bp
SV40 origin of replication
SV40 ori
3199 .. 3334  =  136 bp
SV40 origin of replication
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