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DNA Cloning Service

Binaries

p6o2x35s-LH B483

Ligation of 35s-Xba-xho* into B473p6oM-LH*

12345678910pVS1Sm/SpP2x35sColE1hptIIhptIITE9intLBRBXbaI (303)XhoI (1098)PstI, SbfI, NcoI, SpeIBamHI, SmaI, NsiI, EcoRIHindIII, SalI, AflII, BglIIApaI, KpnI, PmeIAatII (3189)BsiWI, NheIB483p6o2x35s-LH10,037 bp
Promoter 1
2x35s
Gene 1
hptII
Features
monocot
Other markers
Sm/Sp
Size
10,037 bp, circular
Record date
22-JAN-2016

Construct

  • P2x35s–hptII · TE9 · selection cassette, 314–2,638
  • MCS · 15 unique sites · at 2,648, 2,798
  • SfiI ×2 · flanks the MCS · at 2,679, 2,743
  • 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
XbaIT^CTAGA303between LB and P2x35s
XhoI (PaeR7I)C^TCGAG1,098between P2x35s and hptII
PstICTGCA^G2,648between TE9 and RB
SbfI (SdaI)CCTGCA^GG2,648between TE9 and RB
NcoIC^CATGG2,674between TE9 and RB
SpeIA^CTAGT2,687between TE9 and RB
BamHIG^GATCC2,693between TE9 and RB
SmaI (XmaI, TspMI)CCC^GGG2,701between TE9 and RB
NsiI (Mph1103I)ATGCA^T2,709between TE9 and RB
EcoRIG^AATTC2,711between TE9 and RB
HindIIIA^AGCTT2,717between TE9 and RB
SalIG^TCGAC2,729between TE9 and RB
AflII (BspTI)C^TTAAG2,739between TE9 and RB
BglIIA^GATCT2,749between TE9 and RB
ApaI (PspOMI)GGGCC^C2,760between TE9 and RB
KpnI (Acc65I)GGTAC^C2,766between TE9 and RB
PmeI (MssI)GTTT^AAAC2,798between TE9 and RB
AatII (ZraI)GACGT^C3,189between RB and ColE1
BsiWI (Pfl23II)C^GTACG7,403in pVS1
NheI (BmtI)G^CTAGC7,519in pVS1

Cut twice: SphI (2,654, 2,923), NdeI (4,304, 6,063), ClaI (2,724, 7,599), EcoRV (966, 3,310), SfiI (2,679, 2,743), FseI (5,070, 7,175), MfeI (14, 1,792), AgeI (8,013, 8,321), StuI (2,012, 2,737), SacII (1,486, 2,316), BstEII (1,406, 8,884), BsmBI (4,482, 5,535), SapI (4,242, 8,139).

No site: SacI, MluI, 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+
P2x35sPromoter misc_feature3141,098785+
hptIISelection marker CDS1,1161,685570+
intSite misc_feature1,6861,874189+
hptIISelection marker CDS1,8752,330456+
TE9Terminator misc_feature2,3482,638291+
RBT-DNA border misc_feature2,8142,83724+
ColE1Origin / backbone misc_feature3,3724,069698+
pVS1Origin / backbone misc_feature4,6188,3233,706+
Sm/SpSelection marker CDS8,5079,5171,011+

PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite

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