Sticky ends from different DraIII sites may not be compatible.
PsiI (5052) 1 site
TTATAAAATATT
AsiSI (4479) 1 site
GCGATCGCCGCTAGCG
PvuI (4479) 1 site
CGATCGGCTAGC
SmaI (4353) 1 site
CCCGGGGGGCCC
SmaI can be used at 37°C for brief incubations.
TspMI (4351) 1 site
CCCGGGGGGCCC
XmaI (4351) 1 site
CCCGGGGGGCCC
Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
NruI (4136) 1 site
TCGCGAAGCGCT
AcuI (3825) 1 site
CTGAAG(N)14NNGACTTC(N)14
Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI (3693) 1 site
CAGNNNCTGGTCNNNGAC
Sticky ends from different AlwNI sites may not be compatible.
BssSαI (3450) 1 site
CACGAGGTGCTC
PciI (3277) 1 site
ACATGTTGTACA
PciI is inhibited by nonionic detergents.
BspQI (3161) 1 site
GCTCTTCNCGAGAAGNNNN
Sticky ends from different BspQI sites may not be compatible.
SapI (3161) 1 site
GCTCTTCNCGAGAAGNNNN
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.
TatI (3081) 1 site
WGTACWWCATGW
BstZ17I (3048) 1 site
GTATACCATATG
PflFI (3022) 1 site
GACNNNGTCCTGNNNCAG
The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible.
Tth111I (3022) 1 site
GACNNNGTCCTGNNNCAG
The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible.
BlpI (80) 1 site
GCTNAGCCGANTCG
Sticky ends from different BlpI sites may not be compatible.
PaeR7I (158) 1 site
CTCGAGGAGCTC
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI (158) 1 site
VCTCGAGBBGAGCTCV
XhoI (158) 1 site
CTCGAGGAGCTC
EagI (166) 1 site
CGGCCGGCCGGC
NotI (166) 1 site
GCGGCCGCCGCCGGCG
HindIII (173) 1 site
AAGCTTTTCGAA
SalI (179) 1 site
GTCGACCAGCTG
Eco53kI (188) 1 site
GAGCTCCTCGAG
SacI (190) 1 site
GAGCTCCTCGAG
EcoRI (192) 1 site
GAATTCCTTAAG
BamHI (198) 1 site
GGATCCCCTAGG
After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
EcoRV (206) 1 site
GATATCCTATAG
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
NcoI (212) 1 site
CCATGGGGTACC
Acc65I (234) 1 site
GGTACCCCATGG
KpnI (238) 1 site
GGTACCCCATGG
BglII (241) 1 site
AGATCTTCTAGA
BstBI (268) 1 site
TTCGAAAAGCTT
NdeI (346) 1 site
CATATGGTATAC
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
XbaI (384) 1 site
TCTAGAAGATCT
SgrAI (495) 1 site
CRCCGGYGGYGGCCRC
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SphI (651) 1 site
GCATGCCGTACG
BstAPI (859) 1 site
GCANNNNNTGCCGTNNNNNACG
Sticky ends from different BstAPI sites may not be compatible.
MluI (1176) 1 site
ACGCGTTGCGCA
BclI (1190) 1 site
TGATCAACTAGT
* Blocked by Dam methylation.BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII (1357) 1 site
GGTNACCCCANTGG
Sticky ends from different BstEII sites may not be compatible.BstEII is typically used at 60°C, but is 50% active at 37°C.
NmeAIII (1382) 1 site
GCCGAG(N)18-19NNCGGCTC(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).
PspOMI (1383) 1 site
GGGCCCCCCGGG
ApaI (1387) 1 site
GGGCCCCCCGGG
ApaI can be used between 25°C and 37°C.
BssHII (1587) 1 site
GCGCGCCGCGCG
BssHII is typically used at 50°C, but is 75% active at 37°C.
HpaI (1682) 1 site
GTTAACCAATTG
PshAI (2021) 1 site
GACNNNNGTCCTGNNNNCAG
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BglI (2240) 1 site
GCCNNNNNGGCCGGNNNNNCCG
Sticky ends from different BglI sites may not be compatible.
FspI (2258) 1 site
TGCGCAACGCGT
FspAI (2258) 1 site
RTGCGCAYYACGCGTR
PpuMI (2283) 1 site
RGGWCCYYCCWGGR
Sticky ends from different PpuMI sites may not be compatible.
lacI 826 .. 1908 = 1083 bp 360 amino acids = 38.6 kDa Product: lac repressor 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).
lacI 826 .. 1908 = 1083 bp 360 amino acids = 38.6 kDa Product: lac repressor 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).
KanR 4048 .. 4863 = 816 bp 271 amino acids = 31.0 kDa Product: aminoglycoside phosphotransferase confers resistance to kanamycin
KanR 4048 .. 4863 = 816 bp 271 amino acids = 31.0 kDa Product: aminoglycoside phosphotransferase confers resistance to kanamycin
ori 3338 .. 3926 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori 3338 .. 3926 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori 4956 .. 5411 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori 4956 .. 5411 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
rop 2717 .. 2908 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein
rop 2717 .. 2908 = 192 bp 63 amino acids = 7.2 kDa Product: Rop protein
lacI promoter 748 .. 825 = 78 bp
lacI promoter 748 .. 825 = 78 bp
MCS 158 .. 217 = 60 bp multiple cloning site
MCS 158 .. 217 = 60 bp multiple cloning site
T7 terminator 26 .. 73 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator 26 .. 73 = 48 bp transcription terminator for bacteriophage T7 RNA polymerase
S-Tag 249 .. 293 = 45 bp 15 amino acids = 1.7 kDa Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag 249 .. 293 = 45 bp 15 amino acids = 1.7 kDa Product: affinity and epitope tag derived from pancreatic ribonuclease A
lac operator 392 .. 416 = 25 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 392 .. 416 = 25 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).
T7 promoter 417 .. 435 = 19 bp promoter for bacteriophage T7 RNA polymerase
T7 promoter 417 .. 435 = 19 bp promoter for bacteriophage T7 RNA polymerase
6xHis 140 .. 157 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag
6xHis 140 .. 157 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag
thrombin site 300 .. 317 = 18 bp 6 amino acids = 627.8 Da Product: thrombin recognition and cleavage site
thrombin site 300 .. 317 = 18 bp 6 amino acids = 627.8 Da Product: thrombin recognition and cleavage site
6xHis 327 .. 344 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag
6xHis 327 .. 344 = 18 bp 6 amino acids = 840.9 Da Product: 6xHis affinity tag
enterokinase site 219 .. 233 = 15 bp 5 amino acids = 606.5 Da Product: enterokinase recognition and cleavage site
enterokinase site 219 .. 233 = 15 bp 5 amino acids = 606.5 Da Product: enterokinase recognition and cleavage site
RBS 356 .. 361 = 6 bp ribosome binding site
RBS 356 .. 361 = 6 bp ribosome binding site
ATG 345 .. 347 = 3 bp 1 amino acid = 149.2 Da Product: start codon
ATG 345 .. 347 = 3 bp 1 amino acid = 149.2 Da Product: start codon
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