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GAANTRY P helper vector (V009967#)

Basic Vector Information

Basic Vector Information
Vector Name GAANTRY P helper Antibiotic Resistance Ampicillin
Length 6419 bp Type Cloning vector
Source Collier R, Thomson JG, Thilmony R.

GAANTRY P helper vector Vector Map

GAANTRY P helper6419 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300f1 oriM13 fwdM13 fwdlac promoterParATP901-1T3 transcriptional terminatorAmpR promoterlacIT7 terminatorM13 revlac operatorlac promoterCAP binding siteColE1 origin of replicationAmpRAmpR promoter

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6419 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P helper, complete sequence.
ACCESSION   MG687275
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6419
                     /organism="Cloning vector GAANTRY P helper"
                     /mol_type="other DNA"
                     /db_xref="taxon:2201080"
     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"
     regulatory      661..778
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    669..690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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)."
     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
                     /regulatory_class="terminator"
                     /note="T3 transcriptional terminator"
     promoter        2969..3073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3074..4165
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI repressor"
                     /label=lacI
                     /protein_id="AWK49533.1"
                     /translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
                     SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
                     SSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
                     SPRALADSLMQLARQVSRLESGQ"
     gene            3074..4165
                     /gene="lacI"
                     /label=lacI
     regulatory      4166..4212
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      4166..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    4294..4310
                     /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(4318..4348)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4363..4384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4672..5260)
                     /direction=LEFT
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     CDS             complement(5431..6291)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin"
                     /protein_id="AWK49534.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5431..6291)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(6292..6396)
                     /gene="bla"
                     /label=AmpR promoter

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