pTagBFP-C

Vector for fusing TagBFP to the N-terminus of a partner protein.

Sequence Author: Evrogen

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PciI (4649) EcoO109I (3829) RsrII (3247) BsrDI (2964) PflFI - Tth111I (2849) FspI (2833) PluTI (2734) SfoI (2732) NarI (2731) KasI (2730) EagI (2637) BspDI * - ClaI * (2571) BseRI (2549) SfiI (2506) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) BsrGI (644) BbsI (902) BsmBI - Esp3I (1033) PshAI (1234) BspEI (1306) BglII (1315) PaeR7I - XhoI (1319) Eco53kI (1324) SacI (1326) EcoRI (1335) PstI (1344) SalI (1345) Acc65I (1351) KpnI (1355) SacII (1358) PspOMI (1359) TspMI - XmaI (1362) ApaI (1363) SmaI (1364) BamHI (1366) XbaI * (1378) BclI * (1388) MfeI (1481) HpaI (1494) MluI (1617) CsiI - SexAI * (2320) pTagBFP-C 4707 bp
PciI  (4649)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoO109I  (3829)
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.
RsrII  (3247)
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  (2964)
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  (2849)
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  (2849)
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.
FspI  (2833)
1 site
T G C G C A A C G C G T
PluTI  (2734)
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  (2732)
1 site
G G C G C C C C G C G G
NarI  (2731)
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  (2730)
1 site
G G C G C C C C G C G G
EagI  (2637)
1 site
C G G C C G G C C G G C
BspDI  (2571)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2571)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BseRI  (2549)
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.
SfiI  (2506)
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.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
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.
SnaBI  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
BsrGI  (644)
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.
BbsI  (902)
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.
BsmBI  (1033)
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  (1033)
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.
PshAI  (1234)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BspEI  (1306)
1 site
T C C G G A A G G C C T
BglII  (1315)
1 site
A G A T C T T C T A G A
PaeR7I  (1319)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1319)
1 site
C T C G A G G A G C T C
Eco53kI  (1324)
1 site
G A G C T C C T C G A G
SacI  (1326)
1 site
G A G C T C C T C G A G
EcoRI  (1335)
1 site
G A A T T C C T T A A G
PstI  (1344)
1 site
C T G C A G G A C G T C
SalI  (1345)
1 site
G T C G A C C A G C T G
Acc65I  (1351)
1 site
G G T A C C C C A T G G
KpnI  (1355)
1 site
G G T A C C C C A T G G
SacII  (1358)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
PspOMI  (1359)
1 site
G G G C C C C C C G G G
TspMI  (1362)
1 site
C C C G G G G G G C C C
XmaI  (1362)
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.
ApaI  (1363)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (1364)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (1366)
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.
XbaI  (1378)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
BclI  (1388)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
MfeI  (1481)
1 site
C A A T T G G T T A A C
HpaI  (1494)
1 site
G T T A A C C A A T T G
MluI  (1617)
1 site
A C G C G T T G C G C A
CsiI  (2320)
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  (2320)
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.
NeoR/KanR
2603 .. 3397  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
2603 .. 3397  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
TagBFP
607 .. 1305  =  699 bp
233 amino acids  =  26.3 kDa
Product: monomeric blue fluorescent protein
mammalian codon-optimized
TagBFP
607 .. 1305  =  699 bp
233 amino acids  =  26.3 kDa
Product: monomeric blue fluorescent protein
mammalian codon-optimized
ori
4005 .. 4593  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4005 .. 4593  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
1623 .. 2078  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
1623 .. 2078  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
2211 .. 2568  =  358 bp
SV40 enhancer and early promoter
SV40 promoter
2211 .. 2568  =  358 bp
SV40 enhancer and early promoter
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
61 .. 364  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
365 .. 568  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
1495 .. 1616  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1495 .. 1616  =  122 bp
SV40 polyadenylation signal
AmpR promoter
2105 .. 2209  =  105 bp
AmpR promoter
2105 .. 2209  =  105 bp
MCS
1306 .. 1371  =  66 bp
multiple cloning site of fluorescent protein plasmids
MCS
1306 .. 1371  =  66 bp
multiple cloning site of fluorescent protein plasmids
HSV TK poly(A) signal
3629 .. 3676  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
3629 .. 3676  =  48 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
2419 .. 2554  =  136 bp
SV40 origin of replication
SV40 ori
2419 .. 2554  =  136 bp
SV40 origin of replication
ORF:  607 .. 1386  =  780 bp
ORF:  259 amino acids  =  28.7 kDa
ORF:  3418 .. 3867  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  2603 .. 3397  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  2775 .. 3161  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  592 .. 1404  =  813 bp
ORF:  270 amino acids  =  28.1 kDa
ORF:  2912 .. 3448  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  3623 .. 3856  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
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