Basic Vector Information
GAANTRY B donor MoroMYB 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 MoroMYB vector Sequence
LOCUS Exported 9281 bp ds-DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector GAANTRY B donor MoroMYB, complete sequence.
ACCESSION MG687282
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9281)
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JOURNAL Unpublished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 9281)
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 9281)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9281
/organism="Cloning vector GAANTRY B donor MoroMYB"
/mol_type="other DNA"
/db_xref="taxon:2201070"
regulatory complement(7..218)
/regulatory_class="terminator"
/note="CaMV 35S terminator"
CDS complement(join(232..772,1568..1697,1795..1912))
/codon_start=1
/product="Moro MYBA"
/label=Moro MYBA
/note="derived from Citrus sinensis"
/protein_id="AWK49560.1"
/translation="MADSLGVRKGAWTGEEDDLLRKCIEKYGEAKWHQVPLRAGLHRCR
KSCRLRWLNYLNPNIKRGEFAADEVDLILRLHKLLGNRWSLIVGRLPGRTANDVKNFWN
THLRKKVDKCCKNNKEMKAKAEKVEKINIIKPQPRTFAKNSQWLKGKGMTSNNLQLGDY
NLGKQSTPSDHHHHHQQQQENETESVWWESFLFGDELDQQGISSSLSRPEEESTTANIF
AEKSPVVTKVTENRVIEAGQSCPTDDFAFDAELWDLLNAK"
regulatory complement(1924..3163)
/regulatory_class="promoter"
/note="Arabidopsis HotHead (HTH) promoter"
misc_feature 3224..3278
/label=TP901 attB
/note="TP901 attB"
misc_feature complement(3300..3324)
/label=Agrobacterium T-DNA right border
/note="Agrobacterium T-DNA right border"
promoter 3483..3511
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
misc_feature 3552..3657
/label=ParA MRS
/note="ParA MRS"
CDS 3700..4233
/codon_start=1
/gene="aacC1"
/product="gentamicin acetyltransferase"
/label=aacC1
/note="confers resistance to gentamicin"
/protein_id="AWK49561.1"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
gene 3700..4233
/gene="aacC1"
/label=aacC1
misc_feature 4309..4414
/label=ParA MRS
/note="ParA MRS"
primer_bind complement(4437..4453)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4461..4477
/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(4485..4515)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4530..4551
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4784..5229
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 5230..6651
/codon_start=1
/gene="sacB"
/product="levansucrase"
/label=sacB
/note="confers sensitivity to sucrose"
/protein_id="AWK49562.1"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
gene 5230..6651
/gene="sacB"
/label=sacB
rep_origin complement(6749..7337)
/direction=LEFT
/label=ColE1 origin of replication
/note="ColE1 origin of replication"
misc_feature 7508..8368
/label=A118 attB
/note="A118 attB"
CDS complement(7508..8368)
/codon_start=1
/gene="AmpR"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin"
/protein_id="AWK49563.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene complement(7508..8368)
/gene="AmpR"
/label=AmpR
promoter complement(8369..8473)
/gene="bla"
/label=AmpR promoter
rep_origin complement(8499..8954)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 9096..9112
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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