Basic Vector Information
- Vector Name:
- GAANTRY B donor
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6312 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Collier R, Thomson JG, Thilmony R.
- Promoter:
- sacB
GAANTRY B donor 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 vector Sequence
LOCUS 40924_964 6312 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector GAANTRY B donor, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6312) 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a versatile and robust Agrobacterium-based gene stacking system generates high quality transgenic plants JOURNAL Unpublished REFERENCE 2 (bases 1 to 6312) AUTHORS Thilmony R. TITLE Direct Submission JOURNAL Submitted (15-DEC-2017) Crop Improvement and Genetics research Unit,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 6312) TITLE Direct Submission REFERENCE 4 (bases 1 to 6312) 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 TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (15-DEC-2017) Crop Improvement and Genetics research Unit, USDA-ARS-WRRC, 800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..6312 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(3..458) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature 647..702 /label=A118 attB /note="A118 attB" misc_feature 1040..1094 /label=TP901 attB /note="TP901 attB" misc_feature 1116..1140 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" promoter 1299..1327 /label=Pc promoter /note="class 1 integron promoter" CDS 1516..2046 /codon_start=1 /label=GmR /note="gentamycin acetyltransferase" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" misc_feature 2125..2230 /label=ParA MRS /note="ParA MRS" primer_bind complement(2253..2269) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(2277..2293) /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(2301..2331) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(2346..2367) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2600..3045 /label=sacB promoter /note="sacB promoter and control region" CDS 3046..4464 /codon_start=1 /label=SacB /note="secreted levansucrase that renders bacterial growth sensitive to sucrose" /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" rep_origin complement(4565..5153) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5327..6184) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6185..6289) /label=AmpR promoter
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