LOCUS       40924_49422        8842 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector XNWT_GW, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8842)
  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 8842)
  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 8842)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8842)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Physiol 1, 24 (2010)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-DEC-2010) Plant Biology, Carnegie Institution, 260 Panama St, 
            Stanford, CA 94305, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(2..1344)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1603..1883
                     /label=TRP1 promoter
     CDS             1884..2555
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
     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..4006
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_feature    4010..4015
                     /label=mini-ORF
                     /note="mini-ORF"
     protein_bind    4016..4140
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4572..4874)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(5222..5878)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(5879..5981)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(6159..6283)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    6315..6425
                     /note="NubG; N-terminal moiety of ubiquitin (ubi4)"
     CDS             6429..6455
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    6456..6458
                     /label=stop codon
                     /note="stop codon"
     terminator      6480..6667
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6691..6707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6715..6731)
                     /label=lac operator
                     /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(6739..6769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6784..6805)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7093..7681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7855..8712)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8713..8817)
                     /label=AmpR promoter
ORIGIN
        1 gaacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
       61 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
      121 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
      181 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
      241 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
      301 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
      361 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
      421 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
      481 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
      541 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
      601 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
      661 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
      721 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
      781 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
      841 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
      901 ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg
      961 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tcaaagcgtt
     1021 tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca
     1081 cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt
     1141 gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg aaaggtagtc
     1201 tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc cttcagcact
     1261 accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat ccttcaatgc
     1321 tatcatttcc tttgatattg gatcatatta agaaaccatt attatcatga cattaaccta
     1381 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1441 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1501 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1561 tgcggcatca gagcagattg tactgagagt gcaccataga tcaacgacat tactatatat
     1621 ataatatagg aagcatttaa tagacagcat cgtaatatat gtgtactttg cagttatgac
     1681 gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca
     1741 atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa
     1801 aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta
     1861 agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat
     1921 tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg
     1981 ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg
     2041 caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact
     2101 tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg
     2161 gcattgatat cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg
     2221 gtttgccagt tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag
     2281 cttcacagaa acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg
     2341 aacttttgga ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct
     2401 tacattttat gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa
     2461 atggcgttat tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta
     2521 acaaaatagc aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat
     2581 ttaagtattg tttgtgcact tgccgatcta tgcggtgtga aataccgcac agatgcgtaa
     2641 ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
     2701 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2761 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2821 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2881 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2941 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3001 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3061 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg
     3121 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3181 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3241 gttttcccag tcacgacgtt gtaaaacgac ggccagtcgt ccaagctttc gcgagctcga
     3301 tatccttttg ttgtttccgg gtgtacaata tggacttcct cttttctggc aaccaaaccc
     3361 atacatcggg attcctataa taccttcgtt ggtctcccta acatgtaggt ggcggagggg
     3421 agatatacaa tagaacagat accagacaag acataatggg ctaaacaaga ctacaccaat
     3481 tacactgcct cattgatggt ggtacataac gaactaatac tgtagcccta gacttgatag
     3541 ccatcatcat atcgaagttt cactaccctt tttccatttg ccatctattg aagtaataat
     3601 aggcgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
     3661 atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga
     3721 cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac
     3781 tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct
     3841 tccagttact tgaatttgaa ataaaaaaag tttgccgctt tgctatcaag tataaataga
     3901 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
     3961 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactcata tgtgaacaag
     4021 tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata tattaaatta
     4081 gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc agtcactatg
     4141 aatcaactac ttagatggta ttagtgacct gtagtcgacc gacagccttc caaatgttct
     4201 tcgggtgatg ctgccaactt agtcgaccga cagccttcca aatgttcttc tcaaacggaa
     4261 tcgtcgtatc cagcctactc gctattgtcc tcaatgccgt attaaatcat aaaaagaaat
     4321 aagaaaaaga ggtgcgagcc tcttttttgt gtgacaaaat aaaaacatct acctattcat
     4381 atacgctagt gtcatagtcc tgaaaatcat ctgcatcaag aacaatttca caactcttat
     4441 acttttctct tacaagtcgt tcggcttcat ctggattttc agcctctata cttactaaac
     4501 gtgataaagt ttctgtaatt tctactgtat cgacctgcag actggctgtg tataagggag
     4561 cctgacattt atattcccca gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg
     4621 tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc catatcggtg
     4681 gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc acgggagact
     4741 ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc gggcgtgtca
     4801 ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt ataggtgtaa
     4861 accttaaact gcatttcacc agcccctgtt ctcgtcagca aaagagccgt tcatttcaat
     4921 aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt cggcacgcag
     4981 acgacgggct tcattctgca tggttgtgct taccagaccg gagatattga catcatatat
     5041 gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg cttcatagca
     5101 tacctctttt tgacatactt cgggtataca tatcagtata tattcttata ccgcaaaaat
     5161 cagcgcgcaa atacgcatac tgttatctgg cttttagtaa gccggatcca cgcggcgttt
     5221 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     5281 ggaagccatc acagacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     5341 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     5401 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     5461 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     5521 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     5581 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     5641 cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa
     5701 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     5761 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     5821 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     5881 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     5941 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa
     6001 agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt
     6061 gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttag
     6121 tcgactacag gtcactaata ccatctaagt agttgattca tagtgactgg atatgttgtg
     6181 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     6241 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggttggtggt ggcggatccg
     6301 gtggaggtgg atcaatgcag attttcgtca agactttgac cggtaaaacc ataacattgg
     6361 aagttgaatc ttccgatacc atcgacaacg ttaagtcgaa aattcaagac aaggaaggaa
     6421 tccctggtta cccatacgac gttccagatt atgcttagct gcagccaagc taattccggg
     6481 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     6541 acaaatttta aagtgactct taggttttaa aacgaaaatt cttgttcttg agtaactctt
     6601 tcctgtaggt caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct
     6661 ctaccggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6781 ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6841 aaacctgtcg tgccagctgg attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6961 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7021 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7441 agcagccact agtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7501 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7741 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7921 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7981 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8041 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8101 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8161 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8221 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8281 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8341 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8401 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8461 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8521 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8581 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8641 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8701 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8761 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8821 atttccccga aaagtgccac ct
//
