pBAD-DEST49

Gateway® destination vector for the regulated expression of N-terminally tagged HP-thioredoxin fusion proteins in bacterial cells.

Sequence Author: Thermo Fisher (Invitrogen)

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NruI (5868) EcoRV (5546) NsiI (5169) SphI (5167) MauBI (5145) PfoI (5077) PflFI - Tth111I (4978) BsaAI (4972) NdeI (4901) PciI (4722) AhdI (3834) BglI (3716) NmeAIII (3687) AseI (3659) FspI (3611) PvuI (3465) BstEII (77) RBS AleI (441) RsrII (448) HP-thioredoxin BspDI * - ClaI * (530) NgoMIV (673) NaeI (675) EagI - NotI (844) PvuII (1064) EcoRI (1164) PasI (1396) BbvCI (1889) SrfI (2035) BmgBI (2069) BstXI (2152) BfuAI - BspMI (2279) PstI (2290) SalI (2292) BclI * (2422) HindIII (2427) BseRI (2448) PmeI (2511) rrnB T2 terminator pBAD-DEST49 6160 bp
NruI  (5868)
1 site
T C G C G A A G C G C T
EcoRV  (5546)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
NsiI  (5169)
1 site
A T G C A T T A C G T A
SphI  (5167)
1 site
G C A T G C C G T A C G
MauBI  (5145)
1 site
C G C G C G C G G C G C G C G C
PfoI  (5077)
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.
PflFI  (4978)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (4978)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsaAI  (4972)
1 site
Y A C G T R R T G C A Y
NdeI  (4901)
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.
PciI  (4722)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (3834)
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.
BglI  (3716)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
NmeAIII  (3687)
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).
AseI  (3659)
1 site
A T T A A T T A A T T A
FspI  (3611)
1 site
T G C G C A A C G C G T
PvuI  (3465)
1 site
C G A T C G G C T A G C
BstEII  (77)
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.
AleI  (441)
1 site
C A C N N N N G T G G T G N N N N C A C
RsrII  (448)
1 site
C G G W C C G G C C W