RBC T&A Cloning Vector (linearized)

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

Sequence Author: RBC Bioscience

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HomePlasmidsTA and GC Cloning VectorsRBC T&A Cloning Vector (linearized)
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) 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) HindIII (39) 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
Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
HindIII  (39)
1 site		 A A G C T T T T C G A A
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 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  (398)
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  (398)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BseYI  (702)
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  (706)
1 site		 C C C A G C G G G T C G
AmpR
1218 .. 2078  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1218 .. 2009  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1218 .. 2078  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2010 .. 2078  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1218 .. 2078  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
459 .. 1047  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
459 .. 1047  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
lacZα
2424 .. 2729  =  306 bp
101 amino acids  =  11.7 kDa
Product: LacZα fragment of β-galactosidase.
lacZα
2424 .. 2729  =  306 bp
101 amino acids  =  11.7 kDa
Product: LacZα fragment of β-galactosidase.
AmpR promoter
2079 .. 2183  =  105 bp
AmpR promoter
2079 .. 2183  =  105 bp
lacZα
4 .. 61  =  58 bp
19 amino acids  =  2.1 kDa
Product: LacZα fragment of β-galactosidase.
lacZα
4 .. 61  =  58 bp
19 amino acids  =  2.1 kDa
Product: LacZα fragment of β-galactosidase.
lac promoter
105 .. 135  =  31 bp
3 segments
   Segment 3:  -10  
   105 .. 111  =  7 bp
promoter for the E. coli lac operon
lac promoter
105 .. 135  =  31 bp
3 segments
   Segment 2:  
   112 .. 129  =  18 bp
promoter for the E. coli lac operon
lac promoter
105 .. 135  =  31 bp
3 segments
   Segment 1:  -35  
   130 .. 135  =  6 bp
promoter for the E. coli lac operon
lac promoter
105 .. 135  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
81 .. 97  =  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-β-D-thiogalactopyranoside (IPTG).
lac operator
81 .. 97  =  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-β-D-thiogalactopyranoside (IPTG).
MCS
4 .. 44  =  41 bp
multiple cloning site
MCS
4 .. 44  =  41 bp
multiple cloning site
MCS
2701 .. 2729  =  29 bp
multiple cloning site
MCS
2701 .. 2729  =  29 bp
multiple cloning site
T7 promoter
2680 .. 2698  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2680 .. 2698  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 rev
57 .. 73  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
57 .. 73  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2657 .. 2673  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2657 .. 2673  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1348 .. 1614  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1218 .. 2078  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2424 .. 2729  =  306 bp
ORF:  101 amino acids  =  11.7 kDa  (no start codon)
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