LOCUS       Exported                7315 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Plant expression vector pDuExP (pDuExAn6), complete sequence.
ACCESSION   EF565883
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7315)
  AUTHORS   Fujikawa Y, Kato N.
  TITLE     Split luciferase complementation assay to study protein-protein 
            interactions in Arabidopsis protoplasts
  JOURNAL   Plant J. 52 (1), 185-195 (2007)
  PUBMED    17662028
REFERENCE   2  (bases 1 to 7315)
  AUTHORS   Kato N.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2007) Dapartment of Biological Sciences, Louisiana
            State University, 226 Life Sciences Building, Baton Rouge, LA 
            70803-1715, USA
REFERENCE   3  (bases 1 to 7315)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7315
                     /organism="Plant expression vector pDuExP (pDuExAn6)"
                     /mol_type="other DNA"
                     /db_xref="taxon:461715"
     misc_feature    9..26
                     /label=6xHis peptide tag
                     /note="6xHis peptide tag"
     CDS             9..26
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             27..713
                     /codon_start=1
                     /product="NhRluc"
                     /label=NhRluc
                     /note="N-terminal fragment of the Renilla luciferase 
                     protein"
                     /protein_id="ABU49619.1"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGG"
     misc_feature    714..744
                     /note="2x(Gly-Gly-Gly-Gly-Ser); linker"
     misc_recomb     750..874
                     /label=attR1
                     /note="attR1"
     protein_bind    750..874
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1021..1123
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1124..1783
                     /codon_start=1
                     /product="CmR"
                     /label=CmR
                     /note="chloramphenicol resistance"
                     /protein_id="ABU49616.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2125..2430
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /protein_id="ABU49618.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_recomb     complement(2465..2595)
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(2471..2595)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2619..2871
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      2625..2877
                     /regulatory_class="terminator"
                     /note="noparine synthase gene terminus"
     primer_bind     complement(2880..2896)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3290..3308)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3326..3342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3350..3366
                     /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(3374..3404)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3419..3440
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3710..4392)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      complement(3728..4316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4487..5347)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(4487..5149)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance gene"
                     /protein_id="ABU49617.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     promoter        complement(5348..5452)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5530..5985)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(5662..5968)
                     /direction=LEFT
                     /label=F1 origin
                     /note="F1 origin"
     primer_bind     6126..6142
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6149..6167
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6198..6207
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        6221..6239
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_recomb     6305..6338
                     /label=loxP
                     /note="loxP"
     protein_bind    6305..6338
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      6371..7203
                     /regulatory_class="promoter"
                     /note="Cauliflower mosaic virus promoter 35S"
     promoter        6858..7203
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    7232..7287
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"