LOCUS       Exported                8919 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector XN21_GW, complete sequence.
ACCESSION   HQ700951
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8919)
  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 8919)
  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 8919)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8919
                     /organism="Expression vector XN21_GW"
                     /mol_type="other DNA"
                     /db_xref="taxon:1027259"
     rep_origin      2..1349
                     /label=2-micron
                     /note="2-micron"
     rep_origin      2..1344
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(965..1012)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1603..1883
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     gene            1782..2604
                     /gene="TRP1"
                     /label=TRP1
     regulatory      1782..1883
                     /regulatory_class="promoter"
                     /gene="TRP1"
     CDS             1884..2558
                     /codon_start=1
                     /gene="TRP1"
                     /product="TRP1"
                     /label=TRP1
                     /protein_id="AED88262.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     regulatory      2559..2604
                     /regulatory_class="terminator"
                     /gene="TRP1"
     rep_origin      complement(2661..3119)
                     /direction=LEFT
                     /label=F1 origin
                     /note="F1 origin"
     rep_origin      complement(2661..3116)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3261..3277
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3302..4008
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     regulatory      3650..4008
                     /regulatory_class="promoter"
                     /note="ADH1"
     misc_feature    4012..4017
                     /label=mini-ORF
                     /note="mini-ORF"
     misc_feature    4018..6285
                     /note="Gateway recombination cassette containing the attR 
                     sites, ccdB and CamR genes"
     misc_recomb     4018..4142
                     /label=attR1
                     /note="attR1"
     protein_bind    4018..4142
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4571..4876)
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /protein_id="AED88263.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(5221..5880)
                     /codon_start=1
                     /product="CamR"
                     /label=CamR
                     /note="chloramphenicol acetyl transferase"
                     /protein_id="AED88264.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(5881..5983)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_recomb     6161..6286
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(6161..6285)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    6287..6316
                     /label=linker
                     /note="linker"
     misc_feature    6317..6427
                     /note="NubG; N-terminal moiety of ubiquitin (ubi4)"
     misc_difference 6353..6355
                     /label=mutation Ile to Gly
                     /note="mutation Ile to Gly"
     misc_feature    6431..6520
                     /label=3x HA-tag
                     /note="3x HA-tag"
     CDS             6431..6520
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    6530..6532
                     /label=stop codon
                     /note="stop codon"
     regulatory      6553..6744
                     /regulatory_class="terminator"
                     /note="ADH1"
     terminator      6553..6740
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6768..6784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6792..6808
                     /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(6816..6846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      7152..7767
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(7170..7758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7929..8789)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /note="beta-lactamase"
                     /protein_id="AED88265.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8790..8894)
                     /gene="bla"
                     /label=AmpR promoter