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Plasmid Files

pRACE (linearized)

Linearized vector for In-Fusion® cloning of products amplified with the SMARTer RACE 5'/3' kit.

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pRACE (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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ApoI - EcoRI (2636) PluTI (2479) SfoI (2477) NarI (2476) KasI (2475) BstAPI (2425) NdeI (2424) PfoI (2286) EcoO109I (2229) AatII (2175) ZraI (2173) SspI (2057) XmnI (1852) ScaI (1733) TsoI (1652) NmeAIII (1401) BsrFI (1333) HindIII (1) BspQI - SapI (244) AflIII - PciI (360) BseYI (664) PspFI (668) AlwNI (776) AhdI (1253) BsaI (1314) BpmI (1323) pRACE 2656 bp
ApoI  (2636)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (2636)
1 site
G A A T T C C T T A A G
PluTI  (2479)
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  (2477)
1 site
G G C G C C C C G C G G
NarI  (2476)
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  (2475)
1 site
G G C G C C C C G C G G
BstAPI  (2425)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
NdeI  (2424)
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.
PfoI  (2286)
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.
EcoO109I  (2229)
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.
AatII  (2175)
1 site
G A C G T C C T G C A G
ZraI  (2173)
1 site
G A C G T C C T G C A G
SspI  (2057)
1 site
A A T A T T T T A T A A
XmnI  (1852)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1733)
1 site
A G T A C T T C A T G A
TsoI  (1652)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
NmeAIII  (1401)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsrFI  (1333)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
HindIII  (1)
1 site
A A G C T T T T C G A A
BspQI  (244)
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  (244)
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.
AflIII  (360)
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  (360)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (664)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PspFI  (668)
1 site
C C C A G C G G G T C G
AlwNI  (776)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (1253)
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  (1314)
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.
BpmI  (1323)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AmpR
1180 .. 2040  =  861 bp
beta-lactamase, confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1180 .. 2040  =  861 bp
beta-lactamase, confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
421 .. 1009  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
421 .. 1009  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
2041 .. 2145  =  105 bp
AmpR promoter
AmpR promoter
2041 .. 2145  =  105 bp
AmpR promoter
lac promoter
67 .. 97  =  31 bp
lac promoter
67 .. 97  =  31 bp
M13 rev
19 .. 35  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
19 .. 35  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
43 .. 59  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG).
lac operator
43 .. 59  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG).
M13 fwd
2619 .. 2635  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2619 .. 2635  =  17 bp
common sequencing primer, one of multiple similar variants
In-Fusion homology to be added to GSP
1 .. 15  =  15 bp
In-Fusion homology to be added to Gene-Specific Primer (GSP)                            
In-Fusion homology to be added to GSP
1 .. 15  =  15 bp
In-Fusion homology to be added to Gene-Specific Primer (GSP)                            
In-Fusion homology to UPM
2642 .. 2656  =  15 bp
In-Fusion homology to Universal Primer Mix (UPM)                            
In-Fusion homology to UPM
2642 .. 2656  =  15 bp
In-Fusion homology to Universal Primer Mix (UPM)                            
ORF:  1310 .. 1576  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2426 .. 2656  =  231 bp
ORF:  77 amino acids  =  9.1 kDa
ORF:  1180 .. 2040  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2386 .. 2655  =  270 bp
ORF:  89 amino acids  =  10.3 kDa  (no start codon)
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