LOCUS       Exported               11502 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRGK 366, complete sequence.
ACCESSION   DQ666282
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11502)
  AUTHORS   Engineer CB, Kranz RG.
  TITLE     Reciprocal leaf and root expression of AtAmt1.1 and root 
            architectural changes in response to nitrogen starvation
  JOURNAL   Plant Physiol. 143 (1), 236-250 (2007)
  PUBMED    17085512
REFERENCE   2  (bases 1 to 11502)
  AUTHORS   Engineer CB, Kranz RG.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-JUN-2006) Biology, Washington University in St. Louis,
            1 Brookings Drive, # 1137, St. Louis, MO 63130, USA
REFERENCE   3  (bases 1 to 11502)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11502
                     /organism="Cloning vector pRGK 366"
                     /mol_type="other DNA"
                     /db_xref="taxon:393482"
     misc_feature    292..316
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    292..315
                     /note="T-DNA; right border"
     misc_feature    331..783
                     /label=LacZ-MCS
                     /note="LacZ-MCS"
     CDS             complement(354..704)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRFKQSTLDLIKDPARPRVPSSNSLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRI
                     WCTLSTICSDAA"
     primer_bind     587..603
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(700..716)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    724..740
                     /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(748..778)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      787..836
                     /regulatory_class="promoter"
                     /note="minimal 35S CaMV promoter element"
     promoter        790..836
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     protein_bind    938..1618
                     /label=GAL4/VP16 binding site
                     /bound_moiety="GAL4/VP16"
     CDS             1381..1614
                     /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      1619..1874
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      1621..1873
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      1882..2283
                     /regulatory_class="promoter"
                     /note="full 35S CaMV promoter"
     promoter        2121..2167
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     gene            2280..2832
                     /gene="BAR"
                     /label=BAR
     CDS             2280..2831
                     /codon_start=1
                     /gene="BAR"
                     /product="BASTA resistance protein"
                     /label=BAR
                     /protein_id="ABG75720.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     misc_feature    2875..3247
                     /label=7S UTR
                     /note="7S UTR"
     protein_bind    3293..3387
                     /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    3294..3388
                     /note="5X UAS; upstream activation sequence; 5 repeats"
     regulatory      3396..3492
                     /regulatory_class="promoter"
                     /note="minimal 35S CaMV promoter element"
     promoter        3448..3494
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             3532..5184
                     /codon_start=1
                     /gene="pspLUC"
                     /product="luciferase"
                     /label=pspLUC
                     /protein_id="ABG75721.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     gene            3532..5184
                     /gene="pspLUC"
                     /label=pspLUC
     misc_feature    5227..6186
                     /label=TML cassette
                     /note="TML cassette"
     protein_bind    6213..6483
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        6524..6570
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    6571..6595
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    6579..6595
                     /note="T-DNA; left border"
     rep_origin      6831..7362
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      6839..7453
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      6845..7238
                     /label=oriV
                     /note="oriV"
     CDS             8742..8933
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    9035..9175
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      9352..9981
                     /label=ori322
                     /note="ori322"
     rep_origin      complement(9361..9949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     gene            10522..11311
                     /gene="spc/str"
                     /label=spc/str
     CDS             10522..11310
                     /codon_start=1
                     /gene="spc/str"
                     /product="spectinomycin/streptomycin resistance"
                     /label=spc/str
                     /protein_id="ABG75722.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"