Basic Vector Information
- Vector Name:
- pRGK335
- Length:
- 11051 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Engineer CB, Fitzsimmons KC, Schmuke JJ, Dotson SB, Kranz RG.
- Promoter:
- minimal CaMV 35S
pRGK335 vector Map
pRGK335 vector Sequence
LOCUS 40924_36963 11051 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pRGK335, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11051)
AUTHORS Engineer CB, Fitzsimmons KC, Schmuke JJ, Dotson SB, Kranz RG.
TITLE Development and evaluation of a Gal4-mediated LUC/GFP/GUS enhancer
trap system in Arabidopsis
JOURNAL BMC Plant Biol. 5, 9 (2005)
PUBMED 15941484
REFERENCE 2 (bases 1 to 11051)
AUTHORS Engineer CB, Fitzsimmons KC, Schmuke J, Dotson S, Kranz RG.
TITLE Direct Submission
JOURNAL Submitted (01-SEP-2004) Biology, Washington University in St. Louis,
1 Brookings Drive, Biology 1137, St. Louis, MO 63130-4899, USA
REFERENCE 3 (bases 1 to 11051)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11051)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "BMC Plant
Biol. 5, 9 (2005)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(01-SEP-2004) Biology, Washington University in St. Louis, 1
Brookings Drive, Biology 1137, St. Louis, MO 63130-4899, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11051
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(292..316)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
promoter 339..385
/label=minimal CaMV 35S promoter
/note="minimal 35S promoter from cauliflower mosaic virus"
CDS 486..926
/codon_start=1
/label=GAL4 DNA binding domain
/note="DNA binding domain of the GAL4 transcriptional
activator"
/translation="MKLLSSIEQACDICRLKKLKCSKKKPKCAKCLKNNWECRYSPKTK
RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
CDS 930..1163
/codon_start=1
/label=VP16 AD
/note="transcriptional activation domain of herpes simplex
virus protein VP16 (Triezenberg et al., 1988; Cousens et
al., 1989)"
/translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
TPHDSAPYGALDTADFEFEQMFTDALGIDEYGG"
terminator 1170..1422
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter 1670..1716
/label=minimal CaMV 35S promoter
/note="minimal 35S promoter from cauliflower mosaic virus"
CDS 1829..2377
/codon_start=1
/label=BlpR
/note="phosphinothricin acetyltransferase"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
protein_bind 2842..2936
/label=5X UAS
/note="five tandem copies of the 'ScaI site' 17-mer
CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS)
that efficiently binds yeast Gal4 (Webster et al., 1988;
Pfeiffer et al., 2010)"
misc_feature 2945..3041
/label=m35S
/note="m35S"
promoter 2997..3043
/label=minimal CaMV 35S promoter
/note="minimal 35S promoter from cauliflower mosaic virus"
CDS 3081..4730
/codon_start=1
/label=luciferase
/note="firefly luciferase"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
misc_feature 4776..5735
/label=TML
/note="TML"
protein_bind 5762..6032
/label=tetracycline response element
/note="contains seven copies of the tetracycline operator
tetO"
promoter 6073..6119
/label=minimal CaMV 35S promoter
/note="minimal 35S promoter from cauliflower mosaic virus"
misc_feature complement(6120..6144)
/label=LB T-DNA repeat
/note="left border repeat from octopine Ach5 T-DNA"
rep_origin complement(6310..7023)
/direction=LEFT
/label=oriV
/note="incP origin of replication"
CDS 8291..8479
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 8584..8724
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(8910..9498)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 10071..10859
/codon_start=1
/product="spectinomycin/streptomycin resistance"
/label=spectinomycin/streptomycin resistance
/note="Spc/Str"
/protein_id="AAY79159.1"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
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