LOCUS       Exported               11700 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRGK336, complete sequence.
ACCESSION   AY739898
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11700)
  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 11700)
  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 11700)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11700
                     /organism="Cloning vector pRGK336"
                     /mol_type="other DNA"
                     /db_xref="taxon:310383"
     misc_feature    1..11700
                     /note="contains full 35S CaMV, GAL4, luciferase, 
                     spectinomycin/streptomycin resistance in bacteria, BASTA 
                     resistance in plants"
     misc_feature    292..316
                     /note="T-DNA; right border"
     regulatory      422..1034
                     /regulatory_class="promoter"
                     /note="e35S; enhanced 35S CaMV promoter"
     promoter        691..1036
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1579..1812
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDTADFEFEQMFTDALGIDEYGG"
     regulatory      1817..2072
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      1819..2071
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      2080..2478
                     /regulatory_class="promoter"
                     /note="full 35S CaMV promoter"
     promoter        2319..2365
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             2478..3029
                     /codon_start=1
                     /product="BASTA resistance protein"
                     /label=BASTA resistance protein
                     /note="BAR"
                     /protein_id="AAY79160.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     misc_feature    3073..3445
                     /label=7S UTR
                     /note="7S UTR"
     protein_bind    3491..3585
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /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    3492..3586
                     /note="5X UAS; upstream activating sequence"
     promoter        3646..3692
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             3730..5382
                     /codon_start=1
                     /product="luciferase"
                     /label=luciferase
                     /note="psp LUC"
                     /protein_id="AAY79161.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    5425..6383
                     /label=TML
                     /note="TML"
     protein_bind    6411..6681
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        6722..6768
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    6769..6793
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    6777..6793
                     /note="T-DNA; left border"
     rep_origin      7029..7560
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      7037..7651
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     misc_feature    7043..7436
                     /label=ori-V
                     /note="ori-V"
     CDS             8940..9131
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    9233..9373
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    9550..10178
                     /label=ori 322
                     /note="ori 322"
     rep_origin      complement(9559..10147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             10720..11508
                     /codon_start=1
                     /product="spectinomycin/streptomycin resistance"
                     /label=spectinomycin/streptomycin resistance
                     /protein_id="AAY79162.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"