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DNA Cloning Service

Binaries

p7oUH-LH B417

Ligation of UbiH-BsmB-Xho* into B414p7o-LH*

123456789pVS1Sm/SpColE1UbiZmbarTE9bombarintmRNALBRBXhoI (653)AarI (1194)MluI, XmnIPstI, SbfI, NcoI, SpeIBamHI, SmaI, NsiI, EcoRIHindIII, SalI, StuI, BglIIKpnI, PmeIAatII, EcoRVBbsI (3673)BsiWI, NheIBstEII (7958)B417p7oUH-LH9,111 bp
Promoter 1
UbiHa
Gene 1
bar
Features
dicots
Other markers
Sm/Sp
Size
9,111 bp, circular
Record date
22-JAN-2016

Construct

  • UbiZm–bar · TE9 · selection cassette, 316–1,712
  • MCS · 14 unique sites · at 1,722, 1,872
  • SfiI ×2 · flanks the MCS · at 1,753, 1,817
  • 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
XhoI (PaeR7I)C^TCGAG653between UbiZm and bar
AarICACCTGC(4/8)1,194in bar
MluIA^CGCGT1,412between bar and TE9
XmnI (PdmI)GAANN^NNTTC1,414between bar and TE9
PstICTGCA^G1,722between TE9 and RB
SbfI (SdaI)CCTGCA^GG1,722between TE9 and RB
NcoIC^CATGG1,748between TE9 and RB
SpeIA^CTAGT1,761between TE9 and RB
BamHIG^GATCC1,767between TE9 and RB
SmaI (XmaI, TspMI)CCC^GGG1,775between TE9 and RB
NsiI (Mph1103I)ATGCA^T1,783between TE9 and RB
EcoRIG^AATTC1,785between TE9 and RB
HindIIIA^AGCTT1,791between TE9 and RB
SalIG^TCGAC1,803between TE9 and RB
StuI (Eco147I)AGG^CCT1,811between TE9 and RB
BglIIA^GATCT1,823between TE9 and RB
KpnI (Acc65I)GGTAC^C1,840between TE9 and RB
PmeI (MssI)GTTT^AAAC1,872between TE9 and RB
AatII (ZraI)GACGT^C2,263between RB and ColE1
EcoRV (Eco32I)GAT^ATC2,384between RB and ColE1
BbsI (BpiI)GAAGAC(2/6)3,673between bom and pVS1
BsiWI (Pfl23II)C^GTACG6,477in pVS1
NheI (BmtI)G^CTAGC6,593in pVS1
BstEII (Eco91I)G^GTNACC7,958in Sm/Sp

Cut twice: XbaI (303, 384), SphI (1,728, 1,997), NdeI (3,378, 5,137), ClaI (1,798, 6,673), SfiI (1,753, 1,817), FseI (4,144, 6,249), MfeI (14, 1,011), AgeI (7,087, 7,395), AflII (320, 1,813), SacII (1,294, 1,311), BsmBI (3,556, 4,609), SapI (3,316, 7,213).

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

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+
UbiZmPromoter misc_feature316651336+
catSite misc_signal4334353+
catSite misc_signal4364383+
catSite misc_signal4694713+
catSite misc_signal4985003+
catSite misc_signal5095113+
catSite misc_signal5305323+
catSite misc_signal5335353+
tataSite misc_signal5385414+
mRNACoding sequence CDS56965183+
barSelection marker CDS671904234+
intSite misc_feature9051,093189+
barSelection marker CDS1,0941,409316+
TE9Terminator misc_feature1,4221,712291+
RBT-DNA border misc_feature1,8881,91124+
ColE1Origin / backbone misc_feature2,4463,143698+
bomOrigin / backbone misc_feature3,1803,439260+
pVS1Origin / backbone misc_feature3,6927,3973,706+
Sm/SpSelection marker CDS7,5818,5911,011+

PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite

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