Basic Vector Information
GAANTRY P donor bar 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 bar vector Sequence
LOCUS Exported 7945 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector GAANTRY P donor bar, complete sequence.
ACCESSION MG687279
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7945)
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JOURNAL Unpublished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 7945)
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 7945)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7945
/organism="Cloning vector GAANTRY P donor bar"
/mol_type="other DNA"
/db_xref="taxon:2201077"
regulatory complement(1..253)
/regulatory_class="terminator"
/note="nos terminator"
terminator 1..253
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(285..836)
/codon_start=1
/gene="bar"
/product="phosphinothricin acetyltransferase"
/label=bar
/note="confers herbicide resistance (BASTA,
phosphinothricin)"
/protein_id="AWK49549.1"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
gene complement(285..836)
/gene="bar"
/label=bar
regulatory complement(887..1439)
/regulatory_class="promoter"
/note="nos promoter"
promoter complement(887..1066)
/label=NOS promoter
/note="nopaline synthase promoter"
primer_bind complement(1473..1489)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1589..1644
/label=A118 attP
/note="A118 attP"
misc_feature complement(1681..1705)
/label=Agrobacterium T-DNA right border
/note="Agrobacterium T-DNA right border"
CDS 2037..2831
/codon_start=1
/gene="nptIII"
/product="aminoglycoside phosphotransferase"
/label=nptIII
/note="confers resistance to kanamycin"
/protein_id="AWK49550.1"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
gene 2037..2831
/gene="nptIII"
/label=nptIII
misc_feature 3023..3128
/label=ParA MRS
/note="ParA MRS"
primer_bind complement(3151..3167)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3175..3191
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(3199..3229)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3244..3265
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3498..3943
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 3944..5365
/codon_start=1
/gene="sacB"
/product="levansucrase"
/label=sacB
/note="confers sensitivity to sucrose"
/protein_id="AWK49551.1"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
gene 3944..5365
/gene="sacB"
/label=sacB
rep_origin complement(5463..6051)
/direction=LEFT
/label=ColE1 origin of replication
/note="ColE1 origin of replication"
CDS complement(6222..7082)
/codon_start=1
/gene="AmpR"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin"
/protein_id="AWK49552.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene complement(6222..7082)
/gene="AmpR"
/label=AmpR
promoter complement(7083..7187)
/gene="bla"
/label=AmpR promoter
rep_origin complement(7213..7668)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 7810..7826
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 7835..7899
/label=TP901 attP
/note="TP901 attP"
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