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DNA Cloning Service

Binaries

p6oU10-LH B433

Ligation of U10-Bsa-Xho* into B413p6o-LH*

12345678910pVS1UBQ10Sm/SpColE1hptIIhptIIintTE9bomintUbi cds 5LBRBSacI (1402)XhoI (1786)MluI (3019)PstI, SbfI, NcoI, SpeIBamHI, SmaI, NsiI, HindIIIAflII, ApaI, KpnI, PmeIAatII (3870)BsiWI, NheIB433p6oU10-LH10,718 bp
Promoter 1
Ubq10
Gene 1
hptII
Features
dicots
Other markers
Sm/Sp
Size
10,718 bp, circular
Record date
22-JAN-2016

Construct

  • UBQ10–hptII · TE9 · selection cassette, 309–3,319
  • MCS · 12 unique sites · at 3,329, 3,479
  • SfiI ×2 · flanks the MCS · at 3,360, 3,424
  • T-DNA borders · LB, RB

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
SacI (Ecl136II)GAGCT^C1,402in UBQ10
XhoI (PaeR7I)C^TCGAG1,786between UBQ10 and hptII
MluIA^CGCGT3,019between hptII and TE9
PstICTGCA^G3,329between TE9 and RB
SbfI (SdaI)CCTGCA^GG3,329between TE9 and RB
NcoIC^CATGG3,355between TE9 and RB
SpeIA^CTAGT3,368between TE9 and RB
BamHIG^GATCC3,374between TE9 and RB
SmaI (XmaI, TspMI)CCC^GGG3,382between TE9 and RB
NsiI (Mph1103I)ATGCA^T3,390between TE9 and RB
HindIIIA^AGCTT3,398between TE9 and RB
AflII (BspTI)C^TTAAG3,420between TE9 and RB
ApaI (PspOMI)GGGCC^C3,441between TE9 and RB
KpnI (Acc65I)GGTAC^C3,447between TE9 and RB
PmeI (MssI)GTTT^AAAC3,479between TE9 and RB
AatII (ZraI)GACGT^C3,870between RB and ColE1
BsiWI (Pfl23II)C^GTACG8,084in pVS1
NheI (BmtI)G^CTAGC8,200in pVS1

Cut twice: EcoRI (969, 3,392), SalI (1,147, 3,410), SphI (3,335, 3,604), NdeI (4,985, 6,744), BglII (1,714, 3,430), ClaI (3,405, 8,280), SfiI (3,360, 3,424), FseI (5,751, 7,856), MfeI (14, 2,480), AgeI (8,694, 9,002), StuI (2,700, 3,418), SacII (2,174, 3,004), BstEII (2,094, 9,565), BbsI (1,589, 5,280), BsmBI (5,163, 6,216).

No site: 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
LBT-DNA border misc_feature12424+
UBQ10Other misc_feature3091,7841,476+
Ubi cds 5Promoter misc_feature1,4131,47866+
intSite misc_feature1,4791,782304+
hptIISelection marker CDS1,8042,373570+
intSite misc_feature2,3742,562189+
hptIISelection marker CDS2,5633,018456+
TE9Terminator misc_feature3,0293,319291+
RBT-DNA border misc_feature3,4953,51824+
ColE1Origin / backbone misc_feature4,0534,750698+
bomOrigin / backbone misc_feature4,7875,046260+
pVS1Origin / backbone misc_feature5,2999,0043,706+
Sm/SpSelection marker CDS9,18810,1981,011+

PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite

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