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DNA Cloning Service

Binaries

p5oU10-2 B638

Ligation of esps-Xho-Mlu* into B435p9oU10-LH*

1234567891011pVS1UBQ10Sm/SpepspsColE1EPSPSintTE9bomcptintUbi cds 5LBRBXmnI (570)SacI (1402)XhoI, PvuIISacII (3224)MluI (3588)PstI, SbfI, NcoI, SpeIBamHI, SmaI, NsiI, HindIIIStuI, AflII, ApaI, KpnIPmeI (4048)AatII (4439)BsiWI, NheIBstEII (10134)B638p5oU10-211,287 bp
Promoter 1
Ubq10
Gene 1
EPSPS
Features
dicots
Other markers
Sm/Sp
Size
11,287 bp, circular
Record date
08-FEB-2016

Construct

  • UBQ10–cpt–EPSPS · TE9 · payload cassette, 309–3,888
  • MCS · 13 unique sites · at 3,898, 4,048
  • SfiI ×2 · flanks the MCS · at 3,929, 3,993
  • 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
XmnI (PdmI)GAANN^NNTTC570in UBQ10
SacI (Ecl136II)GAGCT^C1,402in UBQ10
XhoI (PaeR7I)C^TCGAG1,786between UBQ10 and cpt
PvuIICAG^CTG1,946in cpt
SacII (Cfr42I)CCGC^GG3,224in epsps
MluIA^CGCGT3,588between epsps and TE9
PstICTGCA^G3,898between TE9 and RB
SbfI (SdaI)CCTGCA^GG3,898between TE9 and RB
NcoIC^CATGG3,924between TE9 and RB
SpeIA^CTAGT3,937between TE9 and RB
BamHIG^GATCC3,943between TE9 and RB
SmaI (XmaI, TspMI)CCC^GGG3,951between TE9 and RB
NsiI (Mph1103I)ATGCA^T3,959between TE9 and RB
HindIIIA^AGCTT3,967between TE9 and RB
StuI (Eco147I)AGG^CCT3,987between TE9 and RB
AflII (BspTI)C^TTAAG3,989between TE9 and RB
ApaI (PspOMI)GGGCC^C4,010between TE9 and RB
KpnI (Acc65I)GGTAC^C4,016between TE9 and RB
PmeI (MssI)GTTT^AAAC4,048between TE9 and RB
AatII (ZraI)GACGT^C4,439between RB and ColE1
BsiWI (Pfl23II)C^GTACG8,653in pVS1
NheI (BmtI)G^CTAGC8,769in pVS1
BstEII (Eco91I)G^GTNACC10,134in Sm/Sp

Cut twice: EcoRI (969, 3,961), SalI (1,147, 3,979), SphI (3,904, 4,173), NdeI (5,554, 7,313), BglII (1,714, 3,999), ClaI (3,974, 8,849), SfiI (3,929, 3,993), FseI (6,320, 8,425), MfeI (14, 2,663), AgeI (9,263, 9,571).

No site: AscI, PacI, SwaI, AvrII, BstBI, HpaI, BsrGI, 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_feature1,4131,47664+
intSite misc_feature1,4791,782304+
cptCoding sequence CDS1,8042,022219+
EPSPSCoding sequence CDS2,0232,556534+
intSite misc_feature2,5572,745189+
epspsCoding sequence CDS2,7463,588843+
TE9Terminator misc_feature3,5983,888291+
RBT-DNA border misc_feature4,0644,08724+
ColE1Origin / backbone misc_feature4,6225,319698+
bomOrigin / backbone misc_feature5,3565,615260+
pVS1Origin / backbone misc_feature5,8689,5733,706+
Sm/SpSelection marker CDS9,75710,7671,011+

PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite

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