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DNA Cloning Service

Binaries

p4oUZm-LH B476

Ligation of UbiZm-Bsa* into B471p4oM-LH*

1234567891011pVS1UbiZmahasSm/SpColE1ahasintTE9intLBRBSbfI, SpeI, BamHI, SmaIEcoRI, HindIII, SalI, AflIIKpnI, PmeIAatII, EcoRVBsiWI (9112)BstEII (10593)B476p4oUZm-LH11,746 bp
Promoter 1
UbiZm
Gene 1
AHAS
Features
monocot
Other markers
Sm/Sp
Size
11,746 bp, circular
Record date
22-JAN-2016

Construct

  • MCS · 10 unique sites · at 4,357, 4,507
  • SfiI ×2 · flanks the MCS · at 4,388, 4,452
  • 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
SbfI (SdaI)CCTGCA^GG4,357between TE9 and RB
SpeIA^CTAGT4,396between TE9 and RB
BamHIG^GATCC4,402between TE9 and RB
SmaI (XmaI, TspMI)CCC^GGG4,410between TE9 and RB
EcoRIG^AATTC4,420between TE9 and RB
HindIIIA^AGCTT4,426between TE9 and RB
SalIG^TCGAC4,438between TE9 and RB
AflII (BspTI)C^TTAAG4,448between TE9 and RB
KpnI (Acc65I)GGTAC^C4,475between TE9 and RB
PmeI (MssI)GTTT^AAAC4,507between TE9 and RB
AatII (ZraI)GACGT^C4,898between RB and ColE1
EcoRV (Eco32I)GAT^ATC5,019between RB and ColE1
BsiWI (Pfl23II)C^GTACG9,112in pVS1
BstEII (Eco91I)G^GTNACC10,593in Sm/Sp

Cut twice: SphI (4,363, 4,632), XhoI (1,009, 1,820), NcoI (1,356, 4,383), NheI (1,429, 9,228), ApaI (1,369, 4,469), SfiI (4,388, 4,452), FseI (6,779, 8,884), AgeI (9,722, 10,030), StuI (2,738, 4,446), NsiI (1,587, 4,418), SapI (5,951, 9,848).

No site: SacI, MluI, AscI, PacI, SwaI, AvrII, BstBI, HpaI, BsrGI, SacII, PvuII, XmnI, 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+
UbiZmCoding sequence CDS3101,8171,508+
intSite misc_feature1,2971,816520+
ahasCoding sequence CDS1,8382,476639+
intSite CDS2,4772,665189+
ahasCoding sequence CDS2,6664,0391,374+
TE9Terminator misc_feature4,0574,347291+
RBT-DNA border misc_feature4,5234,54624+
ColE1Origin / backbone misc_feature5,0815,778698+
pVS1Origin / backbone misc_feature6,32710,0323,706+
Sm/SpSelection marker CDS10,21611,2261,011+

PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite

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