pRGK335 vector (V003744)

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

pRGK33511051 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000RB T-DNA repeatminimal CaMV 35S promoterGAL4 DNA binding domainVP16 ADNOS terminatorminimal CaMV 35S promoterBlpR5X UASm35SluciferaseTMLtetracycline response elementminimal CaMV 35S promoterLB T-DNA repeatoriVropbomoriSpc/Str

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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