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GAANTRY B donor MoroMYB vector (V009977#)

Basic Vector Information

Basic Vector Information
Vector Name GAANTRY B donor MoroMYB Antibiotic Resistance Ampicillin
Length 9281 bp Type Cloning vector
Source Collier R, Thomson JG, Thilmony R.

GAANTRY B donor MoroMYB vector Vector Map

GAANTRY B donor MoroMYB9281 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200CaMV 35S terminatorMoro MYBAArabidopsis HotHead (HTH) promoterTP901 attBAgrobacterium T-DNA right borderPc promoterParA MRSaacC1ParA MRSM13 revlac operatorlac promoterCAP binding sitesacB promotersacBColE1 origin of replicationA118 attBAmpR promoterf1 oriM13 fwd

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

Copy Sequence

Download GeneBank File(.gb)

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  JOURNAL   Unpublished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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