Resources
Plasmid Files

pFRT/lacZeo

Flp recombination target site (FRT) vector, with an SV40 enhancer and promoter, for generating a Flp-In™ host cell line. See also pFRT/lacZeo2.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pFRT lacZeo.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
Download Free Trial Get SnapGene Viewer


BspQI - SapI (8092) AhdI (7087) BsaI (7021) ScaI (6606) XmnI (6487) SwaI (6075) ApaI (5617) PspOMI (5613) BamHI (5338) MfeI (5194) EagI (4138) FseI (4083) BsaAI (119) PflFI - Tth111I (125) SfiI (557) XbaI (641) HindIII (662) Bsu36I (889) Bpu10I (1170) BspDI - ClaI (1489) EcoRV (1778) BclI * (2011) Eco53kI (2603) SacI (2605) BsiWI (3436) BstBI (3729) TspMI - XmaI (3895) SmaI (3897) SgrAI (3921) BmgBI (3941) pFRT/lacZeo 8106 bp
BspQI  (8092)
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  (8092)
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.
AhdI  (7087)
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.
BsaI  (7021)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
ScaI  (6606)
1 site
A G T A C T T C A T G A
XmnI  (6487)
1 site
G A A N N N N T T C C T T N N N N A A G
SwaI  (6075)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
ApaI  (5617)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (5613)
1 site
G G G C C C C C C G G G
BamHI  (5338)
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.
MfeI  (5194)
1 site
C A A T T G G T T A A C
EagI  (4138)
1 site
C G G C C G G C C G G C
FseI  (4083)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
BsaAI  (119)
1 site
Y A C G T R R T G C A Y
PflFI  (125)
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  (125)
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.
SfiI  (557)
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.
XbaI  (641)
1 site
T C T A G A A G A T C T
HindIII  (662)
1 site
A A G C T T T T C G A A
Bsu36I  (889)
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.
Bpu10I  (1170)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
BspDI  (1489)
1 site
A T C G A T T A G C T A
ClaI  (1489)
1 site
A T C G A T T A G C T A
EcoRV  (1778)
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.
BclI  (2011)
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.
Eco53kI  (2603)
1 site
G A G C T C C T C G A G
SacI  (2605)
1 site
G A G C T C C T C G A G
BsiWI  (3436)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BstBI  (3729)
1 site
T T C G A A A A G C T T
TspMI  (3895)
1 site
C C C G G G G G G C C C
XmaI  (3895)
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  (3897)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SgrAI  (3921)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
BmgBI  (3941)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
lacZ
678 .. 3722  =  3045 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
lacZ
678 .. 3722  =  3045 bp
1015 amino acids  =  115.5 kDa
Product: β-galactosidase
AmpR
6300 .. 7160  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   6300 .. 6368  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6300 .. 7160  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   6369 .. 7160  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
6300 .. 7160  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
7331 .. 7919  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
7331 .. 7919  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
BleoR
3804 .. 4181  =  378 bp
125 amino acids  =  13.9 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
3804 .. 4181  =  378 bp
125 amino acids  =  13.9 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 promoter
303 .. 600  =  298 bp
SV40 enhancer and early promoter
SV40 promoter
303 .. 600  =  298 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
5208 .. 5342  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5208 .. 5342  =  135 bp
SV40 polyadenylation signal
AmpR promoter
6195 .. 6299  =  105 bp
AmpR promoter
6195 .. 6299  =  105 bp
small t intron
4568 .. 4633  =  66 bp
SV40 (simian virus 40) small t antigen intron
small t intron
4568 .. 4633  =  66 bp
SV40 (simian virus 40) small t antigen intron
FRT
614 .. 661  =  48 bp
FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011).
FRT
614 .. 661  =  48 bp
FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011).
ATG
609 .. 611  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
609 .. 611  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
SV40 ori
470 .. 604  =  135 bp
SV40 origin of replication
SV40 ori
470 .. 604  =  135 bp
SV40 origin of replication
ORF:  4712 .. 5185  =  474 bp
ORF:  157 amino acids  =  18.2 kDa
ORF:  537 .. 4181  =  3645 bp
ORF:  1214 amino acids  =  137.6 kDa
ORF:  6300 .. 7160  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  1524 .. 1838  =  315 bp
ORF:  104 amino acids  =  12.0 kDa
ORF:  2949 .. 3185  =  237 bp
ORF:  78 amino acids  =  9.0 kDa
ORF:  3672 .. 4361  =  690 bp
ORF:  229 amino acids  =  24.7 kDa
ORF:  5283 .. 6044  =  762 bp
ORF:  253 amino acids  =  28.2 kDa
ORF:  2588 .. 2899  =  312 bp
ORF:  103 amino acids  =  10.5 kDa
ORF:  3716 .. 4087  =  372 bp
ORF:  123 amino acids  =  13.2 kDa
ORF:  5483 .. 5767  =  285 bp
ORF:  94 amino acids  =  10.5 kDa
ORF:  6764 .. 7030  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2018 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter