Basic Vector Information
GAANTRY P donor nptII vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
GAANTRY P donor nptII vector Sequence
LOCUS 40924_984 8526 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector GAANTRY P donor nptII, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8526)
AUTHORS Collier R, Thomson JG, Thilmony R.
TITLE GAANTRY, a versatile and robust Agrobacterium-based gene stacking
system generates high quality transgenic plants
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8526)
AUTHORS Thilmony R.
TITLE Direct Submission
JOURNAL Submitted (15-DEC-2017) Crop Improvement and Genetics research Unit,
USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE 3 (bases 1 to 8526)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8526)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-DEC-2017) Crop Improvement and Genetics research Unit,
USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..8526
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 1..811
/label=Arabidopsis ubiquitin 10 promoter and 5' intron
/note="Arabidopsis ubiquitin 10 promoter and 5' intron"
/regulatory_class="promoter"
CDS 827..1615
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
terminator 1679..1931
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 2086..2141
/label=A118 attP
/note="A118 attP"
misc_feature 2178..2202
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 2534..3325
/label=KanR
/note="aminoglycoside phosphotransferase"
misc_feature 3520..3625
/label=ParA MRS
/note="ParA MRS"
primer_bind complement(3648..3664)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3672..3688)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(3696..3726)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3741..3762)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3995..4440
/label=sacB promoter
/note="sacB promoter and control region"
CDS 4441..5859
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
rep_origin complement(5960..6548)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6722..7579)
/label=AmpR
/note="beta-lactamase"
promoter complement(7580..7684)
/label=AmpR promoter
rep_origin complement(7710..8165)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 8307..8323
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 8332..8396
/label=TP901 attP
/note="TP901 attP"
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