pDOE-09 vector (V007951)

Basic Vector Information

Vector Name:
pDOE-09
Antibiotic Resistance:
Kanamycin
Length:
11589 bp
Type:
Binary vector
Replication origin:
ori
Source/Author:
Gookin TE, Assmann SM.
Promoter:
MAS

pDOE-09 vector Map

pDOE-0911589 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500killed PmlI (CACcTG)LB T-DNA repeatnon-unique SacIMAS terminatorBAR gene remnantMCS2 KpnI sitemTurquoise2exclusive localization to the medial Golgi cisternae; from Arabidopsis AtXylT/XYLT (beta-1,2-xylosyltransferase, At5g55500); characterized in PubMed ID 12943552; confirmed by Saint-Jore-Dupas et al. (Plant Cell 2006;18;3182-3200)mas1'-2' mannopine synthase promoter; mas2' drives MCS2 ORF; mas1' truncated by 96 nt; enhancer elements, TATA box, and start codon missingUBQ10; Arabidopsis ubiquitin10 promoterpDOE mcs1F with XhoITMV OmegaX-NmVen210MCS1 back endOCS terminatorUBQ10; ubiquitin10 promoterTMV-Omega enhancerMCS3 front endStrep-Tag IIFLAGStrep-Tag IICVenus210 specific sequenceMCS3 back endNOS terminatorremnant from pFGC5941M13 fwdRB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriKanR

pDOE-09 vector Sequence

LOCUS       40924_14950       11589 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pDOE-09, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11589)
  AUTHORS   Gookin TE, Assmann SM.
  TITLE     Significant reduction of BiFC non-specific assembly facilitates in 
            planta assessment of heterotrimeric G-protein interactors
  JOURNAL   Plant J. (2014) In press
  PUBMED    25187041
REFERENCE   2  (bases 1 to 11589)
  AUTHORS   Gookin TE, Assmann SM.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-SEP-2014) Department of Biology, The Pennsylvania 
            State University, 208 Mueller Laboratory, University Park, PA 16802,
            USA
REFERENCE   3  (bases 1 to 11589)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11589)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant J. 
            (2014) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-SEP-2014) Department of Biology, The Pennsylvania State 
            University, 208 Mueller Laboratory, University Park, PA 16802, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    99..104
                     /label=non-unique SacI
                     /note="non-unique SacI"
     terminator      complement(111..363)
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    complement(373..406)
                     /label=BAR gene remnant
                     /note="BAR gene remnant"
     misc_feature    407..412
                     /label=MCS2 KpnI site
                     /note="MCS2 KpnI site"
     CDS             complement(416..1129)
                     /label=mTurquoise2
                     /note="enhanced monomeric variant of CFP (Goedhart et al., 
                     2012)"
     sig_peptide     complement(1130..1234)
                     /gene="XT-Golgi-mTq2"
                     /note="exclusive localization to the medial Golgi
                     cisternae; from Arabidopsis AtXylT/XYLT 
                     (beta-1,2-xylosyltransferase, At5g55500); characterized in 
                     PubMed ID 12943552; confirmed by Saint-Jore-Dupas et al. 
                     (Plant Cell 2006;18;3182-3200)"
     misc_feature    1235..1240
                     /label=MCS2 NruI site
                     /note="MCS2 NruI site"
     regulatory      complement(1238..1618)
                     /note="mas1'-2' mannopine synthase promoter; mas2' drives
                     MCS2 ORF; mas1' truncated by 96 nt; enhancer elements, TATA
                     box, and start codon missing"
                     /regulatory_class="promoter"
     promoter        complement(1238..1618)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     regulatory      1631..2268
                     /note="UBQ10; Arabidopsis ubiquitin10 promoter"
                     /regulatory_class="promoter"
     misc_difference 1784
                     /replace="g"
                     /label=killed PmlI (CACcTG)
                     /note="killed PmlI (CACcTG)"
     misc_feature    1965..2268
                     /label=cryptic intron from Norris et al. 1993
                     /note="cryptic intron from Norris et al. 1993"
     misc_difference 2197
                     /replace="t"
                     /label=killed XbaI (aCTAGA)
                     /note="killed XbaI (aCTAGA)"
     primer_bind     2269..2293
                     /label=pDOE mcs1F with XhoI
                     /note="pDOE mcs1F with XhoI"
     misc_feature    2281..2297
                     /label=XhoSac linker
                     /note="XhoSac linker"
     misc_feature    2308..2362
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             2364..3044
                     /codon_start=1
                     /gene="X-NmVen210"
                     /product="mcs1-NmVenus210 fusion protein"
                     /label=X-NmVen210
                     /protein_id="AIN46637.1"
                     /translation="MGRALGTSGGSGGGSGMVSKGEELFTGVVPILVELDGDVNGHKFS
                     VSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSA
                     MPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYN
                     SHNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQS
                     KLSK"
     gene            2364..3044
                     /gene="X-NmVen210"
                     /label=X-NmVen210
     misc_feature    2364..2411
                     /label=MCS1 front end
                     /note="MCS1 front end"
     misc_feature    2412..3044
                     /gene="X-NmVen210"
                     /label=NmVenus210 specific sequence
                     /note="NmVenus210 specific sequence"
     CDS             2412..2930
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     misc_difference 3030..3032
                     /gene="X-NmVen210"
                     /label=A206K monomerizing mutation
                     /note="A206K monomerizing mutation"
     misc_feature    3045..3066
                     /label=MCS1 back end
                     /note="MCS1 back end"
     terminator      3077..3784
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     regulatory      3805..4442
                     /note="UBQ10; ubiquitin10 promoter"
                     /regulatory_class="promoter"
     misc_difference 3958
                     /replace="g"
                     /label=killed PmlI (CACcTG)
                     /note="killed PmlI (CACcTG)"
     misc_difference 4371
                     /replace="t"
                     /label=killed XbaI (aCTAGA)
                     /note="killed XbaI (aCTAGA)"
     primer_bind     4443..4467
                     /label=pDOE mcs3bF with Bsu36I
                     /note="pDOE mcs3bF with Bsu36I"
     misc_feature    4450..4465
                     /label=Bsu361 linker
                     /note="Bsu361 linker"
     regulatory      4466..4531
                     /label=TMV-Omega enhancer
                     /note="TMV-Omega enhancer"
                     /regulatory_class="enhancer"
     misc_feature    4476..4530
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     misc_feature    4532..4579
                     /gene="X-SFS-CVen210"
                     /label=MCS3 front end
                     /note="MCS3 front end"
     CDS             4580..4603
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             4613..4636
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             4643..4666
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     misc_feature    4679..4768
                     /gene="X-SFS-CVen210"
                     /label=CVenus210 specific sequence
                     /note="CVenus210 specific sequence"
     misc_feature    4769..4787
                     /label=MCS3 back end
                     /note="MCS3 back end"
     terminator      4788..5040
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5050..5066
                     /label=remnant from pFGC5941
                     /note="remnant from pFGC5941"
     primer_bind     complement(5091..5107)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5310..5334
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6635..7261
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             7693..8763
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      8832..9026
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    9370..9510
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9696..10284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10374..11165)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"

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