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GAANTRY B donor eGFP vector (V009979#)

Basic Vector Information

Basic Vector Information
Vector Name GAANTRY B donor eGFP Antibiotic Resistance Ampicillin
Length 8899 bp Type Cloning vector
Source Collier R, Thomson JG, Thilmony R.

GAANTRY B donor eGFP vector Vector Map

CaMV 35S terminator eGFP ubq potato ubiquitin 7 promoter and 5' intron TP901 attB Agrobacterium T-DNA right border Pc promoter aacC1 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 A118 attB

GAANTRY B donor eGFP vector Sequence

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LOCUS       Exported                8899 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY B donor eGFP, complete sequence.
ACCESSION   MG687278
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8899)
  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  JOURNAL   Unpublished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 8899)
  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 8899)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8899
                     /organism="Cloning vector GAANTRY B donor eGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:2201071"
     regulatory      complement(8..219)
                     /regulatory_class="terminator"
                     /note="CaMV 35S terminator"
     CDS             complement(235..954)
                     /codon_start=1
                     /gene="eGFP"
                     /product="enhanced green fluorescent protein"
                     /label=eGFP
                     /note="confers green fluorescence"
                     /protein_id="AWK49544.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMVDKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            complement(235..954)
                     /gene="eGFP"
                     /label=eGFP
     CDS             complement(959..1188)
                     /codon_start=1
                     /gene="ubq"
                     /product="ubiquitin monomer"
                     /label=ubq
                     /note="ubiquitin monomer confers enhanced expression in 
                     translational fusions"
                     /protein_id="AWK49545.1"
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLADYNIQKESTLHLVLRLRGG"
     gene            complement(959..1188)
                     /gene="ubq"
                     /label=ubq
     regulatory      complement(1189..2907)
                     /regulatory_class="promoter"
                     /note="potato ubiquitin 7 promoter and 5' intron"
     protein_bind    2796..2807
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     misc_feature    2925..2979
                     /label=TP901 attB
                     /note="TP901 attB"
     misc_feature    complement(3001..3025)
                     /label=Agrobacterium T-DNA right border
                     /note="Agrobacterium T-DNA right border"
     promoter        3184..3212
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3401..3934
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin acetyltransferase"
                     /label=aacC1
                     /note="confers resistance to gentamicin"
                     /protein_id="AWK49546.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            3401..3934
                     /gene="aacC1"
                     /label=aacC1
     misc_feature    4010..4115
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(4138..4154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4162..4178
                     /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(4186..4216)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4231..4252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4485..4930
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4931..6352
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /note="confers sensitivity to sucrose"
                     /protein_id="AWK49547.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            4931..6352
                     /gene="sacB"
                     /label=sacB
     rep_origin      complement(6450..7038)
                     /direction=LEFT
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     CDS             complement(7209..8069)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin"
                     /protein_id="AWK49548.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7209..8069)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(8070..8174)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8200..8655)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8797..8813
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8844..8899
                     /label=A118 attB
                     /note="A118 attB"

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