Basic Vector Information
GAANTRY B donor GUS 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 B donor GUS vector Sequence
LOCUS Exported 10366 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector GAANTRY B donor GUS, complete sequence.
ACCESSION MG687280
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10366)
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JOURNAL Unpublished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 10366)
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 10366)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..10366
/organism="Cloning vector GAANTRY B donor GUS"
/mol_type="other DNA"
/db_xref="taxon:2201069"
regulatory complement(29..281)
/regulatory_class="terminator"
/note="nos terminator"
terminator 29..281
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(353..2203)
/codon_start=1
/gene="GUS"
/product="beta-glucuronidase"
/label=GUS
/note="NLS added to N-term"
/protein_id="AWK49553.1"
/translation="MAPKKKRKVEDPRMLRPVETPTREIKKLDGLWAFSLDRENCGIDQ
RWWESALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAV
THYGKVWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVIT
DENGKKKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVAN
GDVSVELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLR
VGIRSVAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYR
TSHYPYAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQ
AHLQAIKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVN
VMFCDAHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITE
YGVDTLAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRV
GGNKKGIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQ"
gene complement(353..2203)
/gene="GUS"
/label=GUS
CDS complement(353..2164)
/codon_start=1
/gene="uidA (or gusA) from E. coli"
/product="beta-glucuronidase"
/label=GUS
/translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
CDS complement(2177..2197)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
regulatory complement(2216..4312)
/regulatory_class="promoter"
/note="Pinus radiata ubiquitin promoter and 5' intron"
primer_bind complement(4327..4343)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 4392..4446
/label=TP901 attB
/note="TP901 attB"
misc_feature complement(4468..4492)
/label=Agrobacterium T-DNA right border
/note="Agrobacterium T-DNA right border"
promoter 4651..4679
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
CDS 4868..5401
/codon_start=1
/gene="aacC1"
/product="gentamicin acetyltransferase"
/label=aacC1
/note="confers resistance to gentamicin"
/protein_id="AWK49554.1"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
gene 4868..5401
/gene="aacC1"
/label=aacC1
misc_feature 5477..5582
/label=ParA MRS
/note="ParA MRS"
primer_bind complement(5605..5621)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5629..5645
/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(5653..5683)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5698..5719
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5952..6397
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 6398..7819
/codon_start=1
/gene="sacB"
/product="levansucrase"
/label=sacB
/note="confers sensitivity to sucrose"
/protein_id="AWK49555.1"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
gene 6398..7819
/gene="sacB"
/label=sacB
rep_origin complement(7917..8505)
/direction=LEFT
/label=ColE1 origin of replication
/note="ColE1 origin of replication"
CDS complement(8676..9536)
/codon_start=1
/gene="AmpR"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin"
/protein_id="AWK49556.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene complement(8676..9536)
/gene="AmpR"
/label=AmpR
promoter complement(9537..9641)
/gene="bla"
/label=AmpR promoter
rep_origin complement(9667..10122)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 10264..10280
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 10311..10366
/label=A118 attB
/note="A118 attB"
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