pDNR-LIB

Donor vector for transferring a cloned cDNA into an acceptor vector using Cre recombinase in the Creator™ system.

Sequence Author: Clontech (TaKaRa)

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TaqII (4147) BmtI (4114) NheI (4110) BglII (4082) M13 fwd MluI (4059) BssSI - BssSαI (3831) BciVI (3806) PspFI (3704) BseYI (3700) ApaLI (3690) AlwNI (3595) BspHI (3284) AscI (3170) BfuAI - BspMI (3148) BstZ17I (2851) EarI (2486) StuI * (2481) MslI (2240) BsgI (2159) SalI (45) NdeI (61) TspMI - XmaI (66) SmaI (68) EcoRI (71) PstI - SbfI (93) MfeI (148) NmeAIII (285) PaeR7I - XhoI (311) HindIII (317) XbaI (323) BstXI (336) BmrI (344) PspOMI (346) ApaI (350) stop codons AvrII (384) SpeI (403) Eco53kI (517) SacI (519) ScaI (558) SspI (663) NcoI (670) PasI (740) PflMI * (746) BsmBI - Esp3I (747) BpmI (855) BsrDI (958) BspEI (975) ZraI (1262) AatII (1264) SacII (1645) BsaAI - SnaBI (1973) BsrGI (2038) pDNR-LIB 4161 bp
TaqII  (4147)
1 site
G A C C G A ( N ) 9 N N C T G G C T ( N ) 9

Sticky ends from different TaqII sites may not be compatible.
BmtI  (4114)
1 site
G C T A G C C G A T C G
NheI  (4110)
1 site
G C T A G C C G A T C G
BglII  (4082)
1 site
A G A T C T T C T A G A
MluI  (4059)
1 site
A C G C G T T G C G C A
BssSI  (3831)
1 site
C A C G A G G T G C T C
BssSαI  (3831)
1 site
C A C G A G G T G C T C
BciVI  (3806)
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.
PspFI  (3704)
1 site
C C C A G C G G G T C G
BseYI  (3700)
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.
ApaLI  (3690)
1 site
G T G C A C C A C G T G
AlwNI  (3595)
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.
BspHI  (3284)
1 site
T C A T G A A G T A C T
AscI  (3170)
1 site
G G C G C G C C C C G C G C G G
BfuAI  (3148)
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  (3148)
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.
BstZ17I  (2851)
1 site
G T A T A C C A T A T G
EarI  (2486)
1 site
C T C T T C N G A G A A G N N N N

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
StuI  (2481)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
MslI  (2240)
1 site
C A Y N N N N R T G G T R N N N N Y A C
BsgI  (2159)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
SalI  (45)
1 site
G T C G A C C A G C T G
NdeI  (61)
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.
TspMI  (66)
1 site
C C C G G G G G G C C C
XmaI  (66)
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  (68)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
EcoRI  (71)
1 site
G A A T T C C T T A A G
PstI  (93)
1 site
C T G C A G G A C G T C
SbfI  (93)
1 site
C C T G C A G G G G A C G T C C
MfeI  (148)
1 site
C A A T T G G T T A A C
NmeAIII  (285)
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).
PaeR7I  (311)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (311)
1 site
C T C G A G G A G C T C
HindIII  (317)
1 site
A A G C T T T T C G A A
XbaI  (323)
1 site
T C T A G A A G A T C T
BstXI  (336)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
BmrI  (344)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
PspOMI  (346)
1 site
G G G C C C C C C G G G
ApaI  (350)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (384)
1 site
C C T A G G G G A T C C
SpeI  (403)
1 site
A C T A G T T G A T C A
Eco53kI  (517)
1 site
G A G C T C C T C G A G
SacI  (519)
1 site
G A G C T C C T C G A G
ScaI  (558)
1 site
A G T A C T T C A T G A
SspI  (663)
1 site
A A T A T T T T A T A A
NcoI  (670)
1 site
C C A T G G G G T A C C
PasI  (740)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (746)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BsmBI  (747)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (747)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BpmI  (855)
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.
BsrDI  (958)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
BspEI  (975)
1 site
T C C G G A A G G C C T
ZraI  (1262)
1 site
G A C G T C C T G C A G
AatII  (1264)
1 site
G A C G T C C T G C A G
SacII  (1645)
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.
BsaAI  (1973)
1 site
Y A C G T R R T G C A Y
SnaBI  (1973)
1 site
T A C G T A A T G C A T
BsrGI  (2038)
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.
SacB
1302 .. 2723  =  1422 bp
473 amino acids  =  53.0 kDa
2 segments
   Segment 2:  
   1302 .. 2636  =  1335 bp
   444 amino acids  =  50.0 kDa
Product: secreted levansucrase that renders bacterial growth sensitive to sucrose
negative selection marker
SacB
1302 .. 2723  =  1422 bp
473 amino acids  =  53.0 kDa
2 segments
   Segment 1:  signal peptide  
   2637 .. 2723  =  87 bp
   29 amino acids  =  3.0 kDa
Product: secreted levansucrase that renders bacterial growth sensitive to sucrose
negative selection marker
SacB
1302 .. 2723  =  1422 bp
473 amino acids  =  53.0 kDa
2 segments
Product: secreted levansucrase that renders bacterial growth sensitive to sucrose
negative selection marker
CmR
530 .. 1189  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
530 .. 1189  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
ori
3360 .. 3948  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3360 .. 3948  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
sacB promoter
2724 .. 3169  =  446 bp
sacB promoter and control region
sacB promoter
2724 .. 3169  =  446 bp
sacB promoter and control region
stuffer fragment
100 .. 291  =  192 bp
stuffer fragment
100 .. 291  =  192 bp
lambda t0 terminator
415 .. 509  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
415 .. 509  =  95 bp
transcription terminator from phage lambda
MCS B
292 .. 351  =  60 bp
multiple cloning site
MCS B
292 .. 351  =  60 bp
multiple cloning site
MCS A
45 .. 99  =  55 bp
multiple cloning site
MCS A
45 .. 99  =  55 bp
multiple cloning site
loxP
9 .. 42  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
9 .. 42  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
1218 .. 1251  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
1218 .. 1251  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
T7 promoter
4089 .. 4107  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
4089 .. 4107  =  19 bp
promoter for bacteriophage T7 RNA polymerase
M13 fwd
4065 .. 4081  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
4065 .. 4081  =  17 bp
common sequencing primer, one of multiple similar variants
stop codons
354 .. 364  =  11 bp
stop codons in all three reading frames
stop codons
354 .. 364  =  11 bp
stop codons in all three reading frames
SV40 poly(A) signal
162 .. 296  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
162 .. 296  =  135 bp
SV40 polyadenylation signal
ORF:  385 .. 612  =  228 bp
ORF:  75 amino acids  =  8.2 kDa
ORF:  1302 .. 2723  =  1422 bp
ORF:  473 amino acids  =  53.0 kDa
ORF:  530 .. 1234  =  705 bp
ORF:  234 amino acids  =  27.3 kDa
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