Basic Vector Information
GAANTRY P helper vector Vector Map
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 helper vector Sequence
LOCUS 40924_999 6419 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector GAANTRY P helper, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6419)
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JOURNAL Unpublished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 6419)
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 6419)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6419)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-DEC-2017) Crop Improvement and Genetics research Unit,
USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..6419
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 633..649
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 669..690
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 705..735
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 743..759
/label=lac operator
/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)."
CDS 779..1447
/codon_start=1
/gene="ParA"
/product="ParA"
/label=ParA
/note="small serine site-specific recombinase"
/protein_id="AWK49531.1"
/translation="MATREQQPEGRRLKVARIYLRASTDEQNLERQESLVAATRAAGYY
VAGIYREKASGARADRPELLRMIADLQPGEVVVAEKIDRISRLPLAEAERLVASIRAKG
AKLAVPGVVDLSELAAEANGVAKIVLESVQDMLLKLALQMARDDYEDRRERQRQGVQLA
KAAGRYTGRKRDAGMHDRIIALRSGGSSIAKTAKLVGCSPSQVKRVWAAWNAQQQNKRA
"
gene 779..1447
/gene="ParA"
/label=ParA
CDS 1449..2906
/codon_start=1
/gene="TP901-1"
/product="TP901-1"
/label=TP901-1
/note="large serine site-specific recombinase"
/protein_id="AWK49532.1"
/translation="MTKKVAIYTRVSTTNQAEEGFSIDEQIDRLTKYAEAMGWQVSDTY
TDAGFSGAKLERPAMQRLINDIENKAFDTVLVYKLDRLSRSVRDTLYLVKDVFTKNKID
FISLNESIDTSSAMGSLFLTILSAINEFERENIKERMTMGKLGRAKSGKSMMWTKTAFG
YYHNRKTGILEIVPLQATIVEQIFTDYLSGISLTKLRDKLNESGHIGKDIPWSYRTLRQ
TLDNPVYCGYIKFKDSLFEGMHKPIIPYETYLKVQKELEERQQQTYERNNNPRPFQAKY
MLSGMARCGYCGAPLKIVLGHKRKDGSRTMKYHCANRFPRKTKGITVYNDNKKCDSGTY
DLSNLENTVIDNLIGFQENNDSLLKIINGNNQPILDTSSFKKQISQIDKKIQKNSDLYL
NDFITMDELKDRTDSLQAEKKLLKAKISENKFNDSTDVFELVKTQLGSIPINELSYDNK
KKIVNNLVSKVDVTADNVDIIFKFQLA"
gene 1449..2906
/gene="TP901-1"
/label=TP901-1
regulatory 2907..2955
/label=T3 transcriptional terminator
/note="T3 transcriptional terminator"
/regulatory_class="terminator"
promoter 2969..3073
/label=AmpR promoter
CDS 3083..4162
/label=lacI
/note="lac repressor"
terminator 4165..4212
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(4270..4286)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4294..4310)
/label=lac operator
/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(4318..4348)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4363..4384)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4672..5260)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5434..6291)
/label=AmpR
/note="beta-lactamase"
promoter complement(6292..6396)
/label=AmpR promoter
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