LOCUS       Exported                8643 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Plant expression vector pDuExB2 (pDuExD7), complete sequence.
ACCESSION   EF565885
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8643)
  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 8643)
  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 8643)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8643
                     /organism="Plant expression vector pDuExB2 (pDuExD7)"
                     /mol_type="other DNA"
                     /db_xref="taxon:461714"
     CDS             36..281
                     /codon_start=1
                     /product="ChRluc"
                     /label=ChRluc
                     /note="C-terminal fragment of the Renilla luciferase 
                     protein"
                     /protein_id="ABU49628.1"
                     /translation="KPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFP
                     NTEFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    282..299
                     /note="Cys-Cys-Pro-Gly-Cys-Cys; tetra-Cys tag"
     CDS             282..299
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     terminator      324..576
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      330..582
                     /regulatory_class="terminator"
                     /note="noparine synthase gene terminus"
     primer_bind     complement(585..601)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(906..1565)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_recomb     1594..1627
                     /label=loxP
                     /note="loxP"
     protein_bind    1594..1627
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        1642..2087
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2088..3509
                     /codon_start=1
                     /product="levansucrase"
                     /label=levansucrase
                     /note="SacB"
                     /protein_id="ABU49625.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     promoter        3577..3681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3682..4542
                     /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             3880..4542
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance gene"
                     /protein_id="ABU49626.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     regulatory      4637..5319
                     /regulatory_class="promoter"
                     /note="ColE1 origin"
     rep_origin      4713..5301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5418..5434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5442..5460
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_recomb     5526..5559
                     /label=loxP
                     /note="loxP"
     protein_bind    5526..5559
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      5592..6424
                     /regulatory_class="promoter"
                     /note="Cauliflower mosaic virus promoter 35S"
     promoter        6079..6424
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    6436..6491
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             6508..6753
                     /codon_start=1
                     /product="ChRluc"
                     /label=ChRluc
                     /note="C-terminal fragment of the Renilla luciferase 
                     protein"
                     /protein_id="ABU49629.1"
                     /translation="KPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFP
                     NTEFVKVKGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_recomb     6790..6914
                     /label=attR1
                     /note="attR1"
     protein_bind    6790..6914
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        7061..7163
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7164..7823
                     /codon_start=1
                     /product="CmR"
                     /label=CmR
                     /note="chloramphenicol resistance"
                     /protein_id="ABU49627.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             8165..8470
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             8258..8470
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /protein_id="ABU49630.1"
                     /translation="MVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVI
                     GEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    complement(8505..8635)
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(8511..8635)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"