pSMART FOS v2.0 (linearized)

Linearized and dephosphorylated fosmid vector for transcription-free cloning with inducible amplification of copy number.

Sequence Author: Lucigen

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HomePlasmidsLucigen VectorspSMART FOS v2.0 (linearized)
AhdI (7573) NotI (7561) SP6 promoter SL1 Primer (7513 .. 7533) NspI (7256) BstZ17I (7239) SacII (6602) FseI (6598) BciVI - MreI (6587) XcmI (6397) AanI (6157) AanI (5961) StuI (5909) XbaI (5887) EcoNI (5613) SmaI (5590) TspMI - XmaI (5588) FspAI (5331) BsrGI (5299) SbfI (5062) BlpI (4601) AfeI (4517) MfeI (4092) NdeI (4076) KpnI (3876) AhdI (8) NotI (15) SwaI (50) SR4 Primer (51 .. 71) ScaI (851) AvrII (881) SalI (1416) HpaI (1453) BstEII (1473) SexAI * (1477) PspOMI (2111) ApaI (2115) BsaI (2269) PfoI * (2327) AleI (2540) PvuI (3212) BclI * (3281) BseRI (3385) SnaBI (3452) Acc65I (3872) pSMART® FOS v2.0 7581 bp
AhdI  (7573)
2 sites		 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.
NotI  (7561)
2 sites		 G C G G C C G C C G C C G G C G
NspI  (7256)
1 site		 R C A T G Y Y G T A C R
BstZ17I  (7239)
1 site		 G T A T A C C A T A T G
SacII  (6602)
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.
FseI  (6598)
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.
BciVI  (6587)
1 site		 G T A T C C ( N ) 5 N C A T A G G ( N ) 5
The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
MreI  (6587)
1 site		 C G C C G G C G G C G G C C G C
XcmI  (6397)
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.
AanI  (6157)
2 sites		 T T A T A A A A T A T T
AanI  (5961)
2 sites		 T T A T A A A A T A T T
StuI  (5909)
1 site		 A G G C C T T C C G G A
XbaI  (5887)
1 site		 T C T A G A A G A T C T
EcoNI  (5613)
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.
SmaI  (5590)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
TspMI  (5588)
1 site		 C C C G G G G G G C C C
XmaI  (5588)
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.
FspAI  (5331)
1 site		 R T G C G C A Y Y A C G C G T R
BsrGI  (5299)
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.
SbfI  (5062)
1 site		 C C T G C A G G G G A C G T C C
BlpI  (4601)
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.
AfeI  (4517)
1 site		 A G C G C T T C G C G A
MfeI  (4092)
1 site		 C A A T T G G T T A A C
NdeI  (4076)
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.
KpnI  (3876)
1 site		 G G T A C C C C A T G G
AhdI  (8)
2 sites		 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.
NotI  (15)
2 sites		 G C G G C C G C C G C C G G C G
SwaI  (50)
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.
ScaI  (851)
1 site		 A G T A C T T C A T G A
AvrII  (881)
1 site		 C C T A G G G G A T C C
SalI  (1416)
1 site		 G T C G A C C A G C T G
HpaI  (1453)
1 site		 G T T A A C C A A T T G
BstEII  (1473)
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.
SexAI  (1477)
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.
PspOMI  (2111)
1 site		 G G G C C C C C C G G G
ApaI  (2115)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
BsaI  (2269)
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.
PfoI  (2327)
1 site		 T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
AleI  (2540)
1 site		 C A C N N N N G T G G T G N N N N C A C
PvuI  (3212)
1 site		 C G A T C G G C T A G C
BclI  (3281)
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.
BseRI  (3385)
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.
SnaBI  (3452)
1 site		 T A C G T A A T G C A T
Acc65I  (3872)
1 site		 G G T A C C C C A T G G
SL1 Primer
21-mer  /  57% GC
1 binding site
7513 .. 7533  =  21 annealed bases
Tm  =  59°C
SR4 Primer
21-mer  /  33% GC
1 binding site
51 .. 71  =  21 annealed bases
Tm  =  52°C
sopA
3178 .. 4353  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F plasmid
sopA
3178 .. 4353  =  1176 bp
391 amino acids  =  44.1 kDa
Product: partitioning protein for the bacterial F plasmid
repE
4932 .. 5687  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the bacterial F plasmid
repE
4932 .. 5687  =  756 bp
251 amino acids  =  29.4 kDa
Product: replication initiation protein for the bacterial F plasmid
CmR
221 .. 880  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
221 .. 880  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
oriV
6073 .. 6687  =  615 bp
origin of replication for the bacterial F plasmid
oriV
6073 .. 6687  =  615 bp
origin of replication for the bacterial F plasmid
sopC
1661 .. 2134  =  474 bp
centromere-like partitioning region of the bacterial F plasmid
sopC
1661 .. 2134  =  474 bp
centromere-like partitioning region of the bacterial F plasmid
cos
1002 .. 1400  =  399 bp
lambda cos site; allows packaging into phage lambda particles
cos
1002 .. 1400  =  399 bp
lambda cos site; allows packaging into phage lambda particles
incC
4679 .. 4929  =  251 bp
incompatibility region of the bacterial F plasmid
incC
4679 .. 4929  =  251 bp
incompatibility region of the bacterial F plasmid
ori2
5778 .. 5997  =  220 bp
secondary origin of replication for the bacterial F plasmid; also known as oriS
ori2
5778 .. 5997  =  220 bp
secondary origin of replication for the bacterial F plasmid; also known as oriS
cat promoter
118 .. 220  =  103 bp
promoter of the E. coli cat gene
cat promoter
118 .. 220  =  103 bp
promoter of the E. coli cat gene
rrnB T1 terminator
69 .. 114  =  46 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
69 .. 114  =  46 bp
transcription terminator T1 from the E. coli rrnB gene
tonB terminator
894 .. 925  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
tonB terminator
894 .. 925  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
T7Te terminator
7454 .. 7481  =  28 bp
phage T7 early transcription terminator
T7Te terminator
7454 .. 7481  =  28 bp
phage T7 early transcription terminator
T7 promoter
28 .. 46  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
28 .. 46  =  19 bp
promoter for bacteriophage T7 RNA polymerase
SP6 promoter
7540 .. 7558  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
SP6 promoter
7540 .. 7558  =  19 bp
promoter for bacteriophage SP6 RNA polymerase
sopB
2207 .. 3178  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F plasmid
sopB
2207 .. 3178  =  972 bp
323 amino acids  =  35.4 kDa
Product: partitioning protein for the bacterial F plasmid
ORF:  221 .. 880  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  7205 .. 7549  =  345 bp
ORF:  114 amino acids  =  13.0 kDa
ORF:  2901 .. 3137  =  237 bp
ORF:  78 amino acids  =  8.3 kDa
ORF:  5298 .. 5531  =  234 bp
ORF:  77 amino acids  =  9.1 kDa
ORF:  2428 .. 2826  =  399 bp
ORF:  132 amino acids  =  15.6 kDa
ORF:  3178 .. 4353  =  1176 bp
ORF:  391 amino acids  =  44.1 kDa
ORF:  4932 .. 5687  =  756 bp
ORF:  251 amino acids  =  29.4 kDa
ORF:  2207 .. 3178  =  972 bp
ORF:  323 amino acids  =  35.4 kDa
ORF:  5138 .. 5383  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  5468 .. 5698  =  231 bp
ORF:  76 amino acids  =  8.7 kDa
ORF:  7082 .. 7429  =  348 bp
ORF:  115 amino acids  =  12.9 kDa
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Individual Sequences & Maps

pAviTag C-His Kan
pEZ BAC
pRANGER-BTB-2
pSMART GC HK (linearized)
pAviTag N-His Kan
pEZ BAC (BamHI linearized)
pRANGER-BTB-3
pSMART GC LK (linearized)
pAviTag-C N-His SUMO Kan
pEZ BAC (HpaI linearized)
pRANGER-BTB-5
pSMART HCAmp
pAviTag-N N-His SUMO Kan
pGC Blue (linearized)
pRham C-His Kan
pSMART HCAmp (linearized)
pETite C-His Kan
pInvRecA
pRham N-His Kan
pSMART HCKan
pETite N-His Kan
pJAZZ-OC
pRham N-His SUMO Kan
pSMART HCKan (linearized)
pETite N-His SUMO Kan
pJAZZ-OK
pSMART BAC v2.0
pSMART LCAmp
pEZSeq-Amp
pJW168
pSMART BAC v2.0 (BamHI linearized)
pSMART LCAmp (linearized)
pEZSeq-Amp (linearized)
pME-HA (linearized)
pSMART BAC v2.0 (EcoRI linearized)
pSMART LCKan
pEZSeq-Kan
pNGS FOS (linearized)
pSMART BAC v2.0 (HindIII linearized)
pSMART LCKan (linearized)
pEZSeq-Kan (linearized)
pRANGER-BTB-1
pSMART FOS v2.0 (linearized)

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