LOCUS       Exported               10247 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P donor luc, complete sequence.
ACCESSION   MG687277
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10247)
  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  JOURNAL   Unpublished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 10247)
  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 10247)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10247
                     /organism="Cloning vector GAANTRY P donor luc"
                     /mol_type="other DNA"
                     /db_xref="taxon:2201078"
     protein_bind    51..72
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        87..117
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    125..141
                     /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)."
     primer_bind     149..165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(204..456)
                     /regulatory_class="terminator"
                     /note="nos terminator"
     terminator      204..456
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(511..2163)
                     /codon_start=1
                     /gene="luc"
                     /product="luciferase"
                     /label=luc
                     /note="firefly luciferase reporter"
                     /protein_id="AWK49539.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     gene            complement(511..2163)
                     /gene="luc"
                     /label=luc
     CDS             complement(2170..2400)
                     /codon_start=1
                     /gene="ubq"
                     /product="ubiquitin monomer"
                     /label=ubq
                     /note="potato ubiquitin monomer; translational fusion 
                     confers enhanced expression"
                     /protein_id="AWK49540.1"
                     /translation="MQIFVKTLTGKTITLEVEGSDTIDNVKAEIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLADYNIQKESTLHLVLRLRGGG"
     gene            complement(2170..2400)
                     /gene="ubq"
                     /label=ubq
     regulatory      complement(2401..3779)
                     /regulatory_class="promoter"
                     /note="potato 409S ubiquitin promoter and 5 intron"
     misc_feature    3887..3942
                     /label=A118 attP
                     /note="A118 attP"
     misc_feature    3979..4003
                     /label=Agrobacterium T-DNA right border
                     /note="Agrobacterium T-DNA right border"
     CDS             4335..5129
                     /codon_start=1
                     /gene="nptIII"
                     /product="aminoglycoside phosphotransferase"
                     /label=nptIII
                     /note="confers resistance to kanamycin"
                     /protein_id="AWK49541.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            4335..5129
                     /gene="nptIII"
                     /label=nptIII
     misc_feature    5321..5426
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(5449..5465)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5473..5489
                     /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(5497..5527)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5542..5563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5796..6241
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             6242..7663
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /note="confers sensitivity to sucrose"
                     /protein_id="AWK49542.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            6242..7663
                     /gene="sacB"
                     /label=sacB
     rep_origin      complement(7761..8349)
                     /direction=LEFT
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     CDS             complement(8520..9380)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin"
                     /protein_id="AWK49543.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(8520..9380)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(9381..9485)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9511..9966)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     10108..10124
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    10133..10197
                     /label=TP901 attP
                     /note="TP901 attP"