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GAANTRY P donor TBS vector (V009969#)

Basic Vector Information

Basic Vector Information
Vector Name GAANTRY P donor TBS Antibiotic Resistance Ampicillin
Length 8480 bp Type Cloning vector
Source Collier R, Thomson JG, Thilmony R.

GAANTRY P donor TBS vector Vector Map

GAANTRY P donor TBS8480 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400enhancer blocking insulator; derived from petunia; transformation booster sequence (TBS)A118 attPAgrobacterium T-DNA right bordernptIIIM13 revlac operatorlac promoterCAP binding sitesacB promotersacBColE1 origin of replicationAmpRAmpR promoterf1 oriM13 fwdTP901 attP

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 P donor TBS vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                8480 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P donor TBS, complete sequence.
ACCESSION   MG687281
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8480)
  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  JOURNAL   Unpublished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 8480)
  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 8480)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8480
                     /organism="Cloning vector GAANTRY P donor TBS"
                     /mol_type="other DNA"
                     /db_xref="taxon:2201076"
     regulatory      1..2018
                     /regulatory_class="insulator"
                     /note="enhancer blocking insulator; derived from petunia; 
                     transformation booster sequence (TBS)"
     misc_feature    2118..2173
                     /label=A118 attP
                     /note="A118 attP"
     misc_feature    complement(2210..2234)
                     /label=Agrobacterium T-DNA right border
                     /note="Agrobacterium T-DNA right border"
     CDS             2566..3360
                     /codon_start=1
                     /gene="nptIII"
                     /product="aminoglycoside phosphotransferase"
                     /label=nptIII
                     /note="confers resistance to kanamycin"
                     /protein_id="AWK49557.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            2566..3360
                     /gene="nptIII"
                     /label=nptIII
     primer_bind     complement(3680..3696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3704..3720
                     /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(3728..3758)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3773..3794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4027..4472
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4473..5894
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /note="confers sensitivity to sucrose"
                     /protein_id="AWK49558.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            4473..5894
                     /gene="sacB"
                     /label=sacB
     rep_origin      complement(5992..6580)
                     /direction=LEFT
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     CDS             complement(6751..7611)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin"
                     /protein_id="AWK49559.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6751..7611)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(7612..7716)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7742..8197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8339..8355
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8364..8428
                     /label=TP901 attP
                     /note="TP901 attP"

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