LOCUS       sequence-298398-        9059 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses MBP-beta-catenin-His (human beta-catenin as a fusion 
            protein with MBP and His- tags) in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9059)
  AUTHORS   Gavagan M, Fagnan E, Speltz EB, Zalatan JG
  TITLE     The Scaffold Protein Axin Promotes Signaling Specificity within the 
            Wnt Pathway by Suppressing Competing Kinase Reactions.
  JOURNAL   Cell Syst. 2020 Jun 24;10(6):515-525.e5. doi: 
            10.1016/j.cels.2020.05.002. Epub 2020 Jun 17.
  PUBMED    32553184
REFERENCE   2  (bases 1 to 9059)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cels.2020.05.002"; journalName: "Cell Syst"; date: 
            "2020-06-24- 24"; volume: "10"; issue: "6"; pages: "515-525.e5"
COMMENT     Sequence Label: pEF019 MBP-beta-catenin-His
FEATURES             Location/Qualifiers
     source          1..9059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..80
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             81..1163
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(100..119)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1406..1411,1412..1427,1428..1434)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1442..1458
                     /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     1447..1469
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1528..2628
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      2485..2494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2602..2625
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2677..2688
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2731..2751
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2755..5097
                     /codon_start=1
                     /gene="CTNNB1"
                     /product="protein involved both in cell-cell adhesion as 
                     part of the cadherin protein complex, and in gene 
                     transcription as part of the Wnt signaling pathway"
                     /label=CTNNB1
                     /note="also known as catenin beta-1; related to Drosophila 
                     armadillo"
                     /translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPS
                     LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     CDS             2755..5097
                     /codon_start=1
                     /gene="Homo sapiens CTNNB1"
                     /product="human cadherin-associated protein that also plays
                     a role in the canonical Wnt signaling pathway"
                     /label=beta-catenin
                     /note="also known as catenin beta-1"
                     /translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPS
                     LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     CDS             5107..5124
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5151..5168)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5151..5167)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5160..5182)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5346..5363)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5516..5602
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5694..5721
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5741..5832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5833..5901,5902..6693)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6051..6070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6735..7248)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6930..6951)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7142..7161
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      7359..7947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7848..7867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8101..8118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8133..8273
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8199..8218)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8359..8381
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8375..8566)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcacttcacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg
     1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
     1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     2461 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg
     2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
     2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcaactagt
     2701 ggatctggtg gtggtggcgg ttccatgagc gagaatcttt attttcaggg atccatggct
     2761 actcaagctg atttgatgga gttggacatg gccatggaac cagacagaaa agcggctgtt
     2821 agtcactggc agcaacagtc ttacctggac tctggaatcc attctggtgc cactaccaca
     2881 gctccttctc tgagtggtaa aggcaatcct gaggaagagg atgtggatac ctcccaagtc
     2941 ctgtatgagt gggaacaggg attttctcag tccttcactc aagaacaagt agctgatatt
     3001 gatggacagt atgcaatgac tcgagctcag agggtacgag ctgctatgtt ccctgagaca
     3061 ttagatgagg gcatgcagat cccatctaca cagtttgatg ctgctcatcc cactaatgtc
     3121 cagcgtttgg ctgaaccatc acagatgctg aaacatgcag ttgtaaactt gattaactat
     3181 caagatgatg cagaacttgc cacacgtgca atccctgaac tgacaaaact gctaaatgac
     3241 gaggaccagg tggtggttaa taaggctgca gttatggtcc atcagctttc taaaaaggaa
     3301 gcttccagac acgctatcat gcgttctcct cagatggtgt ctgctattgt acgtaccatg
     3361 cagaatacaa atgatgtaga aacagctcgt tgtaccgctg ggaccttgca taacctttcc
     3421 catcatcgtg agggcttact ggccatcttt aagtctggag gcattcctgc cctggtgaaa
     3481 atgcttggtt caccagtgga ttctgtgttg ttttatgcca ttacaactct ccacaacctt
     3541 ttattacatc aagaaggagc taaaatggca gtgcgtttag ctggtgggct gcagaaaatg
     3601 gttgccttgc tcaacaaaac aaatgttaaa ttcttggcta ttacgacaga ctgccttcaa
     3661 attttagctt atggcaacca agaaagcaag ctcatcatac tggctagtgg tggaccccaa
     3721 gctttagtaa atataatgag gacctatact tacgaaaaac tactgtggac cacaagcaga
     3781 gtgctgaagg tgctatctgt ctgctctagt aataagccgg ctattgtaga agctggtgga
     3841 atgcaagctt taggacttca cctgacagat ccaagtcaac gtcttgttca gaactgtctt
     3901 tggactctca ggaatctttc agatgctgca actaaacagg aagggatgga aggtctcctt
     3961 gggactcttg ttcagcttct gggttcagat gatataaatg tggtcacctg tgcagctgga
     4021 attctttcta acctcacttg caataattat aagaacaaga tgatggtctg ccaagtgggt
     4081 ggtatagagg ctcttgtgcg tactgtcctt cgggctggtg acagggaaga catcactgag
     4141 cctgccatct gtgctcttcg tcatctgacc agccgacacc aagaagcaga gatggcccag
     4201 aatgcagttc gccttcacta tggactacca gttgtggtta agctcttaca cccaccatcc
     4261 cactggcctc tgataaaggc tactgttgga ttgattcgaa atcttgccct ttgtcccgca
     4321 aatcatgcac ctttgcgtga gcagggtgcc attccacgac tagttcagtt gcttgttcgt
     4381 gcacatcagg atacccagcg ccgtacgtcc atgggtggga cacagcagca atttgtggag
     4441 ggggtccgca tggaagaaat agttgaaggt tgtaccggag cccttcacat cctagctcgg
     4501 gatgttcaca accgaattgt tatcagagga ctaaatacca ttccattgtt tgtgcagctg
     4561 ctttattctc ccattgaaaa catccaaaga gtagctgcag gggtcctctg tgaacttgct
     4621 caggacaagg aagctgcaga agctattgaa gctgagggag ccacagctcc tctgacagag
     4681 ttacttcact ctaggaatga aggtgtggcg acatatgcag ctgctgtttt gttccgaatg
     4741 tctgaggaca agccacaaga ttacaagaaa cggctttcag ttgagctgac cagctctctc
     4801 ttcagaacag agccaatggc ttggaatgag actgctgatc ttggacttga tattggtgcc
     4861 cagggagaac cccttggata tcgccaggat gatcctagct atcgttcttt tcactctggt
     4921 ggatatggcc aggatgcctt gggtatggac cccatgatgg aacatgagat gggtggccac
     4981 caccctggtg ctgactatcc agttgatggg ctgccagatc tggggcatgc ccaggacctc
     5041 atggatgggc tgcctccagg tgacagcaat cagctggcct ggtttgatac tgacctgagc
     5101 ggccgccacc atcaccacca ccactagggg ccgcactcga gaagcttggc actggccgtc
     5161 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     5221 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5281 cagttgcgca gcctgaatgg cgaatggcag cttggctgtt ttggcggatg agataagatt
     5341 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
     5401 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     5461 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     5521 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     5581 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     5641 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     5701 catcctgacg gatggccttt ttgcgtttct acaaactctt tttgtttatt tttctaaata
     5761 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5821 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5881 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5941 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6001 agttttcgcc ccgaagaacg ttctccaatg atgagcactt ttaaagttct gctatgtggc
     6061 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6121 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6181 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6241 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6301 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6361 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6421 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6481 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6541 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6601 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6661 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6721 ctttagattg atttaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat
     6781 aagcaaatat ttaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     6841 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6901 aatagcccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6961 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     7021 aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     7081 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     7141 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     7201 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtaa aaggatctag
     7261 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7321 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7381 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7441 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7501 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7561 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7621 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7681 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7741 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7801 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7861 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7921 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7981 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     8041 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     8101 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     8161 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     8221 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     8281 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     8341 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     8401 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgcagcgat tcacagatgt
     8461 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     8521 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     8581 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     8641 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     8701 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     8761 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     8821 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     8881 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     8941 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     9001 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgaaatt
//
