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DNA Cloning Service

Binaries

p6oN-LH B399

Ligation of hpt-Xho-Mlu* into 715p7N-TE9*

123456789pVS1 ORISm/SpColE1hptIIhptIIPnosTE9intXbaI (303)XhoI (607)MluI (1840)PstI, SbfI, NcoI, SpeIBamHI, SmaI, NsiI, EcoRIHindIII, SalI, AflII, BglIIApaI, KpnI, PmeIAatII, EcoRVBbsI (4101)BsiWI (6905)B399p6oN-LH9,539 bp
Promoter 1
Nos
Gene 1
hptII
Features
dicots
Other markers
Sm/Sp
Size
9,539 bp, circular
Record date
27-JAN-2016

Construct

  • Pnos–hptII · TE9 · selection cassette, 309–2,140
  • MCS · 15 unique sites · at 2,150, 2,300
  • SfiI ×2 · flanks the MCS · at 2,181, 2,245

Read out of the GenBank file; compare every vector in the multiple expression cassettes and cloning sites table.

Files

Order

Vectors from the list cost 250 €, Cas9 vectors 400 €, net. Send the vector name to info@dna-cloning.com and we ship on invoice.

Restriction sites

Unique cutterSiteCuts atLocation
XbaIT^CTAGA303between BL and Pnos
XhoI (PaeR7I)C^TCGAG607between Pnos and hptII
MluIA^CGCGT1,840between hptII and TE9
PstICTGCA^G2,150between TE9 and BR
SbfI (SdaI)CCTGCA^GG2,150between TE9 and BR
NcoIC^CATGG2,176between TE9 and BR
SpeIA^CTAGT2,189between TE9 and BR
BamHIG^GATCC2,195between TE9 and BR
SmaI (XmaI, TspMI)CCC^GGG2,203between TE9 and BR
NsiI (Mph1103I)ATGCA^T2,211between TE9 and BR
EcoRIG^AATTC2,213between TE9 and BR
HindIIIA^AGCTT2,219between TE9 and BR
SalIG^TCGAC2,231between TE9 and BR
AflII (BspTI)C^TTAAG2,241between TE9 and BR
BglIIA^GATCT2,251between TE9 and BR
ApaI (PspOMI)GGGCC^C2,262between TE9 and BR
KpnI (Acc65I)GGTAC^C2,268between TE9 and BR
PmeI (MssI)GTTT^AAAC2,300between TE9 and BR
AatII (ZraI)GACGT^C2,691between BR and ColE1
EcoRV (Eco32I)GAT^ATC2,812between BR and ColE1
BbsI (BpiI)GAAGAC(2/6)4,101between ColE1 and pVS1 ORI
BsiWI (Pfl23II)C^GTACG6,905in pVS1 ORI

Cut twice: SphI (2,156, 2,425), NdeI (3,806, 5,565), NheI (503, 7,021), ClaI (2,226, 7,101), SfiI (2,181, 2,245), FseI (4,572, 6,677), MfeI (14, 1,301), AgeI (7,515, 7,823), StuI (1,521, 2,239), XmnI (1,507, 1,842), BstEII (915, 8,386), BsmBI (3,984, 5,037), SapI (3,744, 7,641).

No site: SacI, AscI, PacI, SwaI, AvrII, BstBI, HpaI, BsrGI, PvuII, AarI.

Computed from the GenBank sequence for 50 common enzymes; positions are the cut in the top strand, without Dam/Dcm methylation. Compare vectors in the restriction site finder.

Annotated features

FeatureTypeStartEndLengthStrand
BLSite misc_feature12424+
PnosPromoter misc_feature309606298+
hptIISelection marker CDS6251,194570+
intSite misc_feature1,1951,383189+
hptIISelection marker CDS1,3841,839456+
TE9Terminator misc_feature1,8502,140291+
BRSite misc_feature2,3162,33924+
ColE1Origin / backbone misc_feature2,8743,571698+
pVS1 ORIOrigin / backbone misc_feature4,1207,8253,706+
Sm/SpSelection marker CDS8,0099,0191,011+

PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite

Feature types are inferred from the labels in the GenBank file; the file itself is the reference.