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DNA Cloning Service

Binaries

p9oN-ocs-LH B403

Ligation of pOCS-BM* into B401p9oN-LH*

123456789pVS1 ORISm/SpColE1nptIIonptIIoPnosTE9TocsintXbaI, SacIIXhoI (607)SacI (1135)XmnI (1620)SbfI, NcoI, SpeI, BamHISmaI, EcoRI, HindIII, SalIStuI, AflII, BglII, KpnIPmeI (2278)AatII, EcoRVBsiWI (6883)ClaI (7079)B403p9oN-ocs-LH9,517 bp
Promoter 1
Nos
Gene 1
nptII
Gene 2
MCS
Features
dicots
Other markers
nptIIo, Sm/Sp
Size
9,517 bp, circular
Record date
22-JAN-2016

Construct

  • Pnos–nptIIo · TE9 · selection cassette, 309–1,918
  • MCS · 8 unique sites · at 1,928, 2,009
  • SfiI ×2 · flanks the MCS · at 1,959, 2,223

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 BL and Pnos
SacII (Cfr42I)CCGC^GG421in Pnos
XhoI (PaeR7I)C^TCGAG607between Pnos and nptIIo
SacI (Ecl136II)GAGCT^C1,135in nptIIo
XmnI (PdmI)GAANN^NNTTC1,620between nptIIo and TE9
SbfI (SdaI)CCTGCA^GG1,928between TE9 and Tocs
NcoIC^CATGG1,954between TE9 and Tocs
SpeIA^CTAGT1,967between TE9 and Tocs
BamHIG^GATCC1,973between TE9 and Tocs
SmaI (XmaI, TspMI)CCC^GGG1,981between TE9 and Tocs
EcoRIG^AATTC1,991between TE9 and Tocs
HindIIIA^AGCTT1,997between TE9 and Tocs
SalIG^TCGAC2,009between TE9 and Tocs
StuI (Eco147I)AGG^CCT2,217between Tocs and BR
AflII (BspTI)C^TTAAG2,219between Tocs and BR
BglIIA^GATCT2,229between Tocs and BR
KpnI (Acc65I)GGTAC^C2,246between Tocs and BR
PmeI (MssI)GTTT^AAAC2,278between Tocs and BR
AatII (ZraI)GACGT^C2,669between BR and ColE1
EcoRV (Eco32I)GAT^ATC2,790between BR and ColE1
BsiWI (Pfl23II)C^GTACG6,883in pVS1 ORI
ClaI (BspDI)AT^CGAT7,079in pVS1 ORI

Cut twice: PstI (1,928, 1,989), SphI (1,934, 2,403), NdeI (3,784, 5,543), NheI (503, 6,999), ApaI (708, 2,240), MluI (1,618, 2,003), SfiI (1,959, 2,223), MfeI (14, 1,028), AgeI (7,493, 7,801), BstEII (1,377, 8,364), SapI (3,722, 7,619), AarI (1,098, 1,219).

No site: AscI, PacI, SwaI, AvrII, BstBI, HpaI, NsiI, 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
BLSite misc_feature12424+
PnosPromoter misc_feature309606298+
nptIIoSelection marker CDS625923299+
spliceSite misc_signal9209234+
intSite CDS9241,110187+
spliceSite misc_signal1,1091,1124+
nptIIoSelection marker CDS1,1111,614504+
TE9Terminator misc_feature1,6281,918291+
TocsTerminator misc_feature2,0142,209196+
BRSite misc_feature2,2942,31724+
ColE1Origin / backbone misc_feature2,8523,549698+
pVS1 ORIOrigin / backbone misc_feature4,0987,8033,706+
Sm/SpSelection marker CDS7,9878,9971,011+

PromoterCoding sequenceTerminatorSelection markerOrigin / backboneT-DNA borderSite

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