LOCUS       Exported                7405 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector NX33_GW, complete sequence.
ACCESSION   HQ700953
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7405)
  AUTHORS   Lalonde S, Sero A, Pratelli R, Pilot G, Chen J, Sardi MI, Parsa SA, 
            Kim DY, Acharya BR, Stein EV, Hu HC, Villiers F, Takeda K, Yang Y, 
            Han YS, Schwacke R, Chiang W, Kato N, Loque D, Assmann SM, Kwak JM, 
            Schroeder JI, Rhee SY, Frommer WB.
  TITLE     A Membrane Protein/Signaling Protein Interaction Network for 
            Arabidopsis Version AMPv2
  JOURNAL   Front Physiol 1, 24 (2010)
  PUBMED    21423366
REFERENCE   2  (bases 1 to 7405)
  AUTHORS   Lalonde S, Frommer WB.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-DEC-2010) Plant Biology, Carnegie Institution, 260 
            Panama St, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 7405)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7405
                     /organism="Expression vector NX33_GW"
                     /mol_type="other DNA"
                     /db_xref="taxon:1027258"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      245..606
                     /regulatory_class="promoter"
                     /note="TRP1"
     misc_feature    607..717
                     /note="NubG; N-terminal moiety of ubiquitin (ubi4)"
     misc_difference 643..645
                     /label=mutation Ile to Gly
                     /note="mutation Ile to Gly"
     misc_feature    718..728
                     /label=linker
                     /note="linker"
     misc_feature    727..2994
                     /note="Gateway recombination cassette containing the attR 
                     sites, ccdB and CamR genes"
     misc_recomb     727..851
                     /label=attR1
                     /note="attR1"
     protein_bind    727..851
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1280..1585)
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /protein_id="AED88270.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1930..2589)
                     /codon_start=1
                     /product="CamR"
                     /label=CamR
                     /note="chloramphenicol acetyl transferase"
                     /protein_id="AED88271.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2590..2692)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_recomb     2870..2994
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(2870..2994)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    3008..3097
                     /label=3x HA-tag
                     /note="3x HA-tag"
     CDS             3008..3097
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    3098..3100
                     /label=stop codon
                     /note="stop codon"
     rep_origin      3101..3908
                     /label=CEN4
                     /note="CEN4"
     misc_feature    3616..4778
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      3941..4778
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     gene            complement(4617..5395)
                     /gene="TRP1"
                     /label=TRP1
     CDS             complement(4617..5291)
                     /codon_start=1
                     /gene="TRP1"
                     /product="TRP1"
                     /label=TRP1
                     /protein_id="AED88272.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     regulatory      complement(5292..5395)
                     /regulatory_class="promoter"
                     /gene="TRP1"
     promoter        complement(5292..5393)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        5499..5603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5604..6464
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /note="beta-lactamase"
                     /protein_id="AED88273.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6626..7241
                     /label=ColE1
                     /note="ColE1"
     rep_origin      6635..7223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"