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Plasmid Files

pBudCE4.1

Mammalian expression vector for the simultaneous expression of tagged proteins from the CMV and EF-1α promoters.

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pBudCE4.1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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FspI (4546) BspHI (4239) BstBI (3122) SfiI (3112) BglII (3099) PaeR7I - XhoI (3093) BstXI (3087) KpnI (3080) Acc65I (3076) BstBI (3071) NotI (3063) PvuI (4567) HindIII (664) PstI - SbfI (680) SalI (682) AccI (683) BspDI * - ClaI * (689) AflII (694) ScaI (703) XbaI (707) BamHI (713) NheI (1876) pBudCE4.1 4595 bp
FspI  (4546)
1 site
T G C G C A A C G C G T
BspHI  (4239)
1 site
T C A T G A A G T A C T
BstBI  (3122)
2 sites
T T C G A A A A G C T T
SfiI  (3112)
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.
BglII  (3099)
1 site
A G A T C T T C T A G A
PaeR7I  (3093)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (3093)
1 site
C T C G A G G A G C T C
BstXI  (3087)
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.
KpnI  (3080)
1 site
G G T A C C C C A T G G
Acc65I  (3076)
1 site
G G T A C C C C A T G G
BstBI  (3071)
2 sites
T T C G A A A A G C T T
NotI  (3063)
1 site
G C G G C C G C C G C C G G C G
PvuI  (4567)
1 site
C G A T C G G C T A G C
HindIII  (664)
1 site
A A G C T T T T C G A A
PstI  (680)
1 site
C T G C A G G A C G T C
SbfI  (680)
1 site
C C T G C A G G G G A C G T C C
SalI  (682)
1 site
G T C G A C C A G C T G
AccI  (683)
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.
BspDI  (689)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (689)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
AflII  (694)
1 site
C T T A A G G A A T T C
ScaI  (703)
1 site
A G T A C T T C A T G A
XbaI  (707)
1 site
T C T A G A A G A T C T
BamHI  (713)
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.
NheI  (1876)
1 site
G C T A G C C G A T C G
EF-1α promoter
1889 .. 3051  =  1163 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
1889 .. 3051  =  1163 bp
strong constitutive promoter for human elongation factor EF-1α
ori
3580 .. 4168  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3580 .. 4168  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
CMV enhancer
10 .. 389  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
10 .. 389  =  380 bp
human cytomegalovirus immediate early enhancer
BleoR
1063 .. 1437  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
1063 .. 1437  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
1551 .. 1867  =  317 bp
SV40 enhancer and early promoter
SV40 promoter
1551 .. 1867  =  317 bp
SV40 enhancer and early promoter
bGH poly(A) signal
3224 .. 3448  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
3224 .. 3448  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
390 .. 593  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
390 .. 593  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
812 .. 933  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
812 .. 933  =  122 bp
SV40 polyadenylation signal
EF-1α MCS
3062 .. 3126  =  65 bp
multiple cloning site after the EF-1α promoter
EF-1α MCS
3062 .. 3126  =  65 bp
multiple cloning site after the EF-1α promoter
CMV MCS
664 .. 718  =  55 bp
multiple cloning site after the CMV promoter
CMV MCS
664 .. 718  =  55 bp
multiple cloning site after the CMV promoter
EM7 promoter
1456 .. 1503  =  48 bp
synthetic bacterial promoter
EM7 promoter
1456 .. 1503  =  48 bp
synthetic bacterial promoter
V5 tag
3127 .. 3168  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
3127 .. 3168  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
Myc
719 .. 748  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc proto-oncogene) epitope tag
Myc
719 .. 748  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc proto-oncogene) epitope tag
T7 promoter
638 .. 656  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
638 .. 656  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
764 .. 781  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
764 .. 781  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3178 .. 3195  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
3178 .. 3195  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
EF-1α intron A
2120 .. 3042  =  923 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
2120 .. 3042  =  923 bp
intron upstream of the start codon of human EF-1α
SV40 ori
1565 .. 1700  =  136 bp
SV40 origin of replication
SV40 ori
1565 .. 1700  =  136 bp
SV40 origin of replication
ORF:  1157 .. 1444  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
ORF:  2429 .. 2866  =  438 bp
ORF:  145 amino acids  =  15.8 kDa
ORF:  2508 .. 2741  =  234 bp
ORF:  77 amino acids  =  7.6 kDa
ORF:  1877 .. 2791  =  915 bp
ORF:  304 amino acids  =  33.0 kDa
ORF:  1063 .. 1437  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  2863 .. 3195  =  333 bp
ORF:  110 amino acids  =  12.7 kDa
ORF:  3214 .. 3522  =  309 bp
ORF:  102 amino acids  =  11.3 kDa
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