LOCUS       40924_49412        8919 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector XN21_GW, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8919)
  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..8919
                     /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..4008
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     misc_feature    4012..4017
                     /label=mini-ORF
                     /note="mini-ORF"
     protein_bind    4018..4142
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4574..4876)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(5224..5880)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(5881..5983)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(6161..6285)
                     /label=attR2
                     /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"
     CDS             6431..6520
                     /label=3xHA
                     /note="three tandem HA epitope tags"
     misc_feature    6530..6532
                     /label=stop codon
                     /note="stop codon"
     terminator      6553..6740
                     /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    complement(6792..6808)
                     /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(6816..6846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6861..6882)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7170..7758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7932..8789)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8790..8894)
                     /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 aaattgtaag 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 gcacacgaaa
     3781 ctttttcctt ccttcattca cgcacactac tctctaatga gcaacggtat acggccttcc
     3841 ttccagttac ttgaatttga aataaaaaaa agtttgctgt cttgctatca agtataaata
     3901 gacctgcaat tattaatctt ttgtttcctc gtcattgttc tcgttccctt tcttccttgt
     3961 ttctttttct gcacaatatt tcaagctata ccaagcatac aatcaactca tatgtgaaca
     4021 agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat
     4081 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta
     4141 tgaatcaact acttagatgg tattagtgac ctgtagtcga ccgacagcct tccaaatgtt
     4201 cttcgggtga tgctgccaac ttagtcgacc gacagccttc caaatgttct tctcaaacgg
     4261 aatcgtcgta tccagcctac tcgctattgt cctcaatgcc gtattaaatc ataaaaagaa
     4321 ataagaaaaa gaggtgcgag cctctttttt gtgtgacaaa ataaaaacat ctacctattc
     4381 atatacgcta gtgtcatagt cctgaaaatc atctgcatca agaacaattt cacaactctt
     4441 atacttttct cttacaagtc gttcggcttc atctggattt tcagcctcta tacttactaa
     4501 acgtgataaa gtttctgtaa tttctactgt atcgacctgc agactggctg tgtataaggg
     4561 agcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg tcattttcgc
     4621 ggtggctgag atcagccact tcttccccga taacggagac cggcacactg gccatatcgg
     4681 tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt tcacgggaga
     4741 ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc ccgggcgtgt
     4801 caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt ttataggtgt
     4861 aaaccttaaa ctgcatttca ccagcccctg ttctcgtcag caaaagagcc gttcatttca
     4921 ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg ttcggcacgc
     4981 agacgacggg cttcattctg catggttgtg cttaccagac cggagatatt gacatcatat
     5041 atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc tgcttcatag
     5101 catacctctt tttgacatac ttcgggtata catatcagta tatattctta taccgcaaaa
     5161 atcagcgcgc aaatacgcat actgttatct ggcttttagt aagccggatc cacgcggcgt
     5221 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     5281 atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     5341 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     5401 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     5461 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     5521 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     5581 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     5641 accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg
     5701 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     5761 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     5821 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     5881 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     5941 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
     6001 aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa
     6061 gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt
     6121 agtcgactac aggtcactaa taccatctaa gtagttgatt catagtgact ggatatgttg
     6181 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     6241 tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggttggtg gtggcggatc
     6301 cggtggaggt ggatcaatgc agattttcgt caagactttg accggtaaaa ccggaacatt
     6361 ggaagttgaa tcttccgata ccatcgacaa cgttaagtcg aaaattcaag acaaggaagg
     6421 aatccctggt tacccatacg atgttcctga ctatgcgggc tatccgtatg acgtcccgga
     6481 ctatgcagga tcctatccat atgacgttcc agattacgct gctcagtgct gaggcgcgcc
     6541 acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa
     6601 aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc
     6661 ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagcatga ggtcgctctt
     6721 attgaccaca cctctaccgg catgcatgca agcttggcgt aatcatggtc atagctgttt
     6781 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6841 tgtaaagcct ggggtgccta atgagtgagg taactcacat taattgcgtt gcgctcactg
     6901 cccgctttcc agtcgggaaa cctgtcgtgc cagctggatt aatgaatcgg ccaacgcgcg
     6961 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7021 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7081 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7801 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7861 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7921 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7981 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8041 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8101 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8161 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8221 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     8281 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8341 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8401 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8461 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8521 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     8581 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8641 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8701 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8761 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8821 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8881 ataggggttc cgcgcacatt tccccgaaaa gtgccacct
//
