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a versatile and robust Agrobacterium-based gene stacking system generates high quality transgenic plants JOURNAL Unpublished REFERENCE 2 (bases 1 to 10366) AUTHORS Thilmony R. TITLE Direct Submission JOURNAL Submitted (15-DEC-2017) Crop Improvement and Genetics research Unit,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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