LOCUS       40924_2269        8899 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector NX32_GW, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8899)
  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 8899)
  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 8899)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8899)
  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..8899
                     /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"
     regulatory      2559..2604
                     /gene="TRP1"
                     /regulatory_class="terminator"
     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    4011..4121
                     /note="NubG; N-terminal moiety of ubiquitin (ubi4)"
     misc_difference 4047..4049
                     /label=mutation Ile to Gly
                     /note="mutation Ile to Gly"
     protein_bind    4131..4255
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4687..4989)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(5337..5993)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(5994..6096)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(6274..6398)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6412..6501
                     /label=3xHA
                     /note="three tandem HA epitope tags"
     misc_feature    6511..6513
                     /label=stop codon
                     /note="stop codon"
     terminator      6534..6721
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6748..6764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6772..6788)
                     /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(6796..6826)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6841..6862)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7150..7738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7912..8769)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8770..8874)
                     /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 tcaactcgag atgcagattt
     4021 tcgtcaagac tttgaccggt aaaaccggaa cattggaagt tgaatcttcc gataccatcg
     4081 acaacgttaa gtcgaaaatt caagacaagg aaggaatccc tggtgggtcg acaagtttgt
     4141 acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
     4201 tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca ctatgaatca
     4261 actacttaga tggtattagt gacctgtagt cgaccgacag ccttccaaat gttcttcggg
     4321 tgatgctgcc aacttagtcg accgacagcc ttccaaatgt tcttctcaaa cggaatcgtc
     4381 gtatccagcc tactcgctat tgtcctcaat gccgtattaa atcataaaaa gaaataagaa
     4441 aaagaggtgc gagcctcttt tttgtgtgac aaaataaaaa catctaccta ttcatatacg
     4501 ctagtgtcat agtcctgaaa atcatctgca tcaagaacaa tttcacaact cttatacttt
     4561 tctcttacaa gtcgttcggc ttcatctgga ttttcagcct ctatacttac taaacgtgat
     4621 aaagtttctg taatttctac tgtatcgacc tgcagactgg ctgtgtataa gggagcctga
     4681 catttatatt ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct
     4741 gagatcagcc acttcttccc cgataacgga gaccggcaca ctggccatat cggtggtcat
     4801 catgcgccag ctttcatccc cgatatgcac caccgggtaa agttcacggg agactttatc
     4861 tgacagcaga cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat
     4921 atcactctgt acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt
     4981 aaactgcatt tcaccagccc ctgttctcgt cagcaaaaga gccgttcatt tcaataaacc
     5041 gggcgacctc agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac
     5101 gggcttcatt ctgcatggtt gtgcttacca gaccggagat attgacatca tatatgcctt
     5161 gagcaactga tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct
     5221 ctttttgaca tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg
     5281 cgcaaatacg catactgtta tctggctttt agtaagccgg atccacgcgg cgtttacgcc
     5341 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     5401 ccatcacaga cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     5461 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     5521 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     5581 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     5641 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     5701 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     5761 ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     5821 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     5881 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     5941 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     6001 tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca
     6061 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg
     6121 gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct
     6181 tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttagtcgac
     6241 tacaggtcac taataccatc taagtagttg attcatagtg actggatatg ttgtgtttta
     6301 cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca
     6361 ttttacgttt ctcgttcagc tttcttgtac aaagtggttg gtggtggcgg atacccatac
     6421 gatgttcctg actatgcggg ctatccgtat gacgtcccgg actatgcagg atcctatcca
     6481 tatgacgttc cagattacgc tgctcagtgc tgaggcgcgc cacttctaaa taagcgaatt
     6541 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     6601 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
     6661 aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg
     6721 gcagcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6781 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6841 atgagtgagg taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6901 cctgtcgtgc cagctggatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6961 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7021 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7501 agccactagt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7561 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7681 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7741 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7801 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7861 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8041 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8221 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8761 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8821 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8881 tccccgaaaa gtgccacct
//
