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GAANTRY P donor nptII vector (V009970#)

Basic Vector Information

Basic Vector Information
Vector Name GAANTRY P donor nptII Antibiotic Resistance Ampicillin
Length 8526 bp Type Cloning vector
Source Collier R, Thomson JG, Thilmony R.

GAANTRY P donor nptII vector Vector Map

Arabidopsis ubiquitin 10 promoter and 5' intron nptII nos terminator A118 attP Agrobacterium T-DNA right border nptIII ParA MRS M13 rev lac operator lac promoter CAP binding site sacB promoter sacB ColE1 origin of replication AmpR AmpR promoter f1 ori M13 fwd TP901 attP

GAANTRY P donor nptII vector Sequence

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LOCUS       Exported                8526 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P donor nptII, complete sequence.
ACCESSION   MG687284
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8526)
  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  JOURNAL   Unpublished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 8526)
  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 8526)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8526
                     /organism="Cloning vector GAANTRY P donor nptII"
                     /mol_type="other DNA"
                     /db_xref="taxon:2201079"
     regulatory      1..811
                     /regulatory_class="promoter"
                     /note="Arabidopsis ubiquitin 10 promoter and 5' intron"
     CDS             821..1618
                     /codon_start=1
                     /gene="nptII"
                     /product="aminoglycoside phosphotransferase"
                     /label=nptII
                     /note="confers resistance to kanamycin"
                     /protein_id="AWK49568.1"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            821..1618
                     /gene="nptII"
                     /label=nptII
     regulatory      1679..1931
                     /regulatory_class="terminator"
                     /note="nos terminator"
     terminator      1679..1931
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2086..2141
                     /label=A118 attP
                     /note="A118 attP"
     misc_feature    complement(2178..2202)
                     /label=Agrobacterium T-DNA right border
                     /note="Agrobacterium T-DNA right border"
     CDS             2534..3328
                     /codon_start=1
                     /gene="nptIII"
                     /product="aminoglycoside phosphotransferase"
                     /label=nptIII
                     /note="confers resistance to kanamycin"
                     /protein_id="AWK49569.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            2534..3328
                     /gene="nptIII"
                     /label=nptIII
     misc_feature    3520..3625
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(3648..3664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3672..3688
                     /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(3696..3726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3741..3762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3995..4440
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4441..5862
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /note="confers sensitivity to sucrose"
                     /protein_id="AWK49570.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            4441..5862
                     /gene="sacB"
                     /label=sacB
     rep_origin      complement(5960..6548)
                     /direction=LEFT
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     CDS             complement(6719..7579)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin"
                     /protein_id="AWK49571.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6719..7579)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(7580..7684)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7710..8165)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8307..8323
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8332..8396
                     /label=TP901 attP
                     /note="TP901 attP"

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