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DNA Cloning Service

Binaries

p9oUbiZmF D015

Ligation of UzmF-Bsa* into B475p9oM-LH*

1234567891011pVS1PUbiZmFSm/SpColE1NptIIoNptIIoTE9intLBRBSacI (2841)SbfI, SpeI, BamHI, SmaIHindIII, StuI, AflII, KpnIPmeI (3791)AatII, EcoRVBsiWI (8396)D015p9oUbiZmF11,030 bp
Promoter 1
UbiZm
Gene 1
nptII
Features
monocots
Other markers
NptIIo, Sm/Sp
Size
11,030 bp, circular
Record date
03-AUG-2020

Construct

  • PUbiZmF–NptIIo · TE9 · selection cassette, 316–3,631
  • MCS · 9 unique sites · at 3,641, 3,791
  • SfiI ×2 · flanks the MCS · at 3,672, 3,736
  • 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^C2,841in NptIIo
SbfI (SdaI)CCTGCA^GG3,641between TE9 and RB
SpeIA^CTAGT3,680between TE9 and RB
BamHIG^GATCC3,686between TE9 and RB
SmaI (XmaI, TspMI)CCC^GGG3,694between TE9 and RB
HindIIIA^AGCTT3,710between TE9 and RB
StuI (Eco147I)AGG^CCT3,730between TE9 and RB
AflII (BspTI)C^TTAAG3,732between TE9 and RB
KpnI (Acc65I)GGTAC^C3,759between TE9 and RB
PmeI (MssI)GTTT^AAAC3,791between TE9 and RB
AatII (ZraI)GACGT^C4,182between RB and ColE1
EcoRV (Eco32I)GAT^ATC4,303between RB and ColE1
BsiWI (Pfl23II)C^GTACG8,396in pVS1

Cut twice: EcoRI (1,707, 3,704), SalI (911, 3,722), SphI (3,647, 3,916), XhoI (1,011, 2,313), NcoI (1,357, 3,667), NheI (1,430, 8,512), BglII (1,257, 3,742), SfiI (3,672, 3,736), AgeI (9,006, 9,314), BstEII (3,083, 9,877), BsmBI (5,475, 6,528), SapI (5,235, 9,132), AarI (2,804, 2,925).

No site: MluI, AscI, PacI, SwaI, AvrII, BstBI, HpaI, BsrGI, SacII, PvuII, XmnI.

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+
PUbiZmFPromoter misc_feature3162,3091,994+
NptIIoSelection marker CDS2,3312,629299+
intSite CDS2,6302,816187+
NptIIoSelection marker CDS2,8173,323507+
TE9Terminator misc_feature3,3413,631291+
RBT-DNA border misc_feature3,8073,83024+
ColE1Origin / backbone misc_feature4,3655,062698+
pVS1Origin / backbone misc_feature5,6119,3163,706+
Sm/SpSelection marker CDS9,50010,5101,011+

PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite

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