pMFAPbeta1

Vector expressing a plasma membrane-localized version of the fluorogen-activating protein FAPβ1.

Sequence Author: SpectraGenetics

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PsiI (6085) DraIII (5960) XmnI (5407) ScaI (5288) PvuI (5178) AhdI (4808) SexAI * (4560) BspDI * - ClaI * (4309) BsaBI * (4292) PluTI (4152) SfoI (4150) NarI (4149) KasI (4148) PflFI - Tth111I (4032) BclI * (3808) RsrII (3638) BspQI - SapI (3600) BstBI (3473) PfoI (3377) BssHII - MauBI (1) CMV enhancer SnaBI (365) HindIII (664) BstXI (709) EcoRV (729) HA PspOMI (828) ApaI (832) SfiI (836) HA NheI (926) BmtI (930) EcoNI (1056) BlpI (1178) BglII (1190) AleI (1205) BstEII (1282) BsmBI - Esp3I (1294) TspMI - XmaI (1397) SmaI (1399) SfiI (1704) SacII (1718) SalI (1727) AccI (1728) BsmI (1768) XcmI (1846) NotI (1917) PaeR7I - XhoI (1923) AflIII - PciI (2235) pMFAP β1 6184 bp
PsiI  (6085)
1 site
T T A T A A A A T A T T
DraIII  (5960)
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.
XmnI  (5407)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (5288)
1 site
A G T A C T T C A T G A
PvuI  (5178)
1 site
C G A T C G G C T A G C
AhdI  (4808)
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.
SexAI  (4560)
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.
BspDI  (4309)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (4309)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
BsaBI  (4292)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
PluTI  (4152)
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  (4150)
1 site
G G C G C C C C G C G G
NarI  (4149)
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  (4148)
1 site
G G C G C C C C G C G G
PflFI  (4032)
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  (4032)
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.
BclI  (3808)
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.
RsrII  (3638)
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.
BspQI  (3600)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3600)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
BstBI  (3473)
1 site
T T C G A A A A G C T T
PfoI  (3377)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BssHII  (1)
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.
MauBI  (1)
1 site
C G C G C G C G G C G C G C G C
SnaBI  (365)
1 site
T A C G T A A T G C A T
HindIII  (664)
1 site
A A G C T T T T C G A A
BstXI  (709)
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.
EcoRV  (729)
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.
PspOMI  (828)
1 site
G G G C C C C C C G G G
ApaI  (832)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SfiI  (836)
2 sites
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.
NheI  (926)
1 site
G C T A G C C G A T C G
BmtI  (930)
1 site
G C T A G C C G A T C G
EcoNI  (1056)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BlpI  (1178)
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.
BglII  (1190)
1 site
A G A T C T T C T A G A
AleI  (1205)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (1282)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BsmBI  (1294)
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  (1294)
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.
TspMI  (1397)
1 site
C C C G G G G G G C C C
XmaI  (1397)
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  (1399)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SfiI  (1704)
2 sites
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.
SacII  (1718)
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.
SalI  (1727)
1 site
G T C G A C C A G C T G
AccI  (1728)
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.
BsmI  (1768)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
XcmI  (1846)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
NotI  (1917)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (1923)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1923)
1 site
C T C G A G G A G C T C
AflIII  (2235)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (2235)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
Igκ leader
737 .. 799  =  63 bp
21 amino acids  =  2.4 kDa
Product: leader sequence from mouse immunoglobulin κ light chain
Igκ leader
737 .. 799  =  63 bp
21 amino acids  =  2.4 kDa
Product: leader sequence from mouse immunoglobulin κ light chain
HA
800 .. 826  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
800 .. 826  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
842 .. 868  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
HA
842 .. 868  =  27 bp
9 amino acids  =  1.1 kDa
Product: HA (human influenza hemagglutinin) epitope tag
FAPβ1
926 .. 1696  =  771 bp
257 amino acids  =  26.4 kDa
Product: fluorogen-activating protein composed of the HL1.0.1-TO1 single-chain antibody fragment (Szent-Gyorgyi et al., 2008)
binds thiazole orange derivatives to produce green fluorescence
FAPβ1
926 .. 1696  =  771 bp
257 amino acids  =  26.4 kDa
Product: fluorogen-activating protein composed of the HL1.0.1-TO1 single-chain antibody fragment (Szent-Gyorgyi et al., 2008)
binds thiazole orange derivatives to produce green fluorescence
Myc
1733 .. 1762  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc proto-oncogene) epitope tag
Myc
1733 .. 1762  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc proto-oncogene) epitope tag
PDGFRβ TM domain
1766 .. 1912  =  147 bp
49 amino acids  =  5.4 kDa
Product: transmembrane domain from platelet derived growth factor receptor beta
PDGFRβ TM domain
1766 .. 1912  =  147 bp
49 amino acids  =  5.4 kDa
Product: transmembrane domain from platelet derived growth factor receptor beta
AmpR
4735 .. 5595  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4735 .. 5526  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4735 .. 5595  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5527 .. 5595  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4735 .. 5595  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
3492 .. 4280  =  789 bp
262 amino acids  =  28.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
NeoR/KanR
3492 .. 4280  =  789 bp
262 amino acids  =  28.7 kDa
Product: aminoglycoside phosphotransferase
confers resistance to neomycin, kanamycin, and G418 (Geneticin®)
ori
2296 .. 2884  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2296 .. 2884  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
5727 .. 6182  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
5727 .. 6182  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
10 .. 389  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
10 .. 389  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
4315 .. 4675  =  361 bp
SV40 enhancer and early promoter
SV40 promoter
4315 .. 4675  =  361 bp
SV40 enhancer and early promoter
bGH poly(A) signal
1940 .. 2164  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
1940 .. 2164  =  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
AmpR promoter
5596 .. 5700  =  105 bp
AmpR promoter
5596 .. 5700  =  105 bp
HSV TK poly(A) signal
3213 .. 3260  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
HSV TK poly(A) signal
3213 .. 3260  =  48 bp
herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)
T7 promoter
638 .. 656  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
638 .. 656  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SV40 ori
4329 .. 4467  =  139 bp
SV40 origin of replication
SV40 ori
4329 .. 4467  =  139 bp
SV40 origin of replication
ORF:  737 .. 1915  =  1179 bp
ORF:  392 amino acids  =  40.7 kDa
ORF:  4865 .. 5131  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3033 .. 3266  =  234 bp
ORF:  77 amino acids  =  8.6 kDa
ORF:  3441 .. 3971  =  531 bp
ORF:  176 amino acids  =  19.6 kDa
ORF:  3728 .. 4108  =  381 bp
ORF:  126 amino acids  =  14.3 kDa
ORF:  1651 .. 1890  =  240 bp
ORF:  79 amino acids  =  8.8 kDa
ORF:  1930 .. 2238  =  309 bp
ORF:  102 amino acids  =  11.3 kDa
ORF:  3022 .. 3471  =  450 bp
ORF:  149 amino acids  =  16.3 kDa
ORF:  4735 .. 5595  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  600 .. 848  =  249 bp
ORF:  82 amino acids  =  9.0 kDa
ORF:  3492 .. 4280  =  789 bp
ORF:  262 amino acids  =  28.7 kDa
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Individual Sequences & Maps

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