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RBC T&A Cloning Vector (linearized)

Linearized vector with 3'-T overhangs for TA cloning of PCR products with blue/white screening.

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RBC T&A Cloning Vector (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  RBC Bioscience
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EcoO109I (2267) AatII (2213) ZraI (2211) SspI (2095) XmnI (1890) ScaI (1771) TsoI (1690) NmeAIII (1439) BsrFI (1371) BpmI (1361) BsaI (1352) AhdI (1291) AlwNI (814) PfoI (2324) NdeI (2462) BstAPI (2463) KasI (2513) NarI (2514) SfoI (2515) PluTI (2517) Eco53kI (2703) BanII - SacI (2705) Acc65I (2707) AvaI - BsoBI - KpnI - TspMI - XmaI (2711) BmeT110I (2712) SmaI (2713) ApoI - EcoRI (2718) End (2730) Start (1) BsaBI * (6) BamHI (7) XbaI (15) SalI (21) AccI (22) HincII (23) PstI - SbfI (31) BfuAI - BspMI (34) SphI (37) BspQI - SapI (282) AflIII - PciI (398) BseYI (702) PspFI (706) RBC T&A Cloning Vector 2729 bp
EcoO109I  (2267)
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  (2213)
1 site
G A C G T C C T G C A G
ZraI  (2211)
1 site
G A C G T C C T G C A G
SspI  (2095)
1 site
A A T A T T T T A T A A
XmnI  (1890)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1771)
1 site
A G T A C T T C A T G A
TsoI  (1690)
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  (1439)
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  (1371)
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.
BpmI  (1361)
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.
BsaI  (1352)
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.
AhdI  (1291)
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.
AlwNI  (814)
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.
PfoI  (2324)
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.
NdeI  (2462)
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.
BstAPI  (2463)
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.
KasI  (2513)
1 site
G G C G C C C C G C G G
NarI  (2514)
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.
SfoI  (2515)
1 site
G G C G C C C C G C G G
PluTI  (2517)
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.
Eco53kI  (2703)
1 site
G A G C T C C T C G A G
BanII  (2705)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (2705)
1 site
G A G C T C C T C G A G
Acc65I  (2707)
1 site
G G T A C C C C A T G G
AvaI  (2711)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2711)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
KpnI  (2711)
1 site
G G T A C C C C A T G G
TspMI  (2711)
1 site
C C C G G G G G G C C C
XmaI  (2711)
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.
BmeT110I  (2712)
1 site
C Y C G R G G R G C Y C
SmaI  (2713)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
ApoI  (2718)
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  (2718)
1 site
G A A T T C C T T A A G
End  (2730)
0 sites
Start  (1)
0 sites
BsaBI  (6)
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.
BamHI  (7)
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  (15)
1 site
T C T A G A A G A T C T
SalI  (21)
1 site
G T C G A C C A G C T G
AccI  (22)
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.
HincII  (23)
1 site
G T Y R A C C A R Y T G
PstI  (31)
1 site
C T G C A G G A C G T C
SbfI  (31)
1 site
C C T G C A G G G G A C G T C C
BfuAI  (34)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (34)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SphI  (37)
1 site
G C A T G C C G T A C G
BspQI  (282)
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  (282)
1 site
G C T C T T C N C G A G A A G N N