LOCUS       sequence-298397-       10852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses MBP-beta-catenin-His (human beta-catenin as a fusion 
            protein with MBP and His- tags) and GST_CK1-alpha in E.coli to 
            produce primed MBP-beta-catenin-His.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10852)
  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 10852)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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: pMG051 MBP-beta-catenin-His, GST-CK1-alpha
FEATURES             Location/Qualifiers
     source          1..10852
                     /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"
     RBS             5154..5176
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5199..5849
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
     primer_bind     5810..5832
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     primer_bind     complement(6944..6961)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6944..6960)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6953..6975)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7139..7156)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      7309..7395
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7487..7514
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        7534..7625
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7626..7694,7695..8486)
                     /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(7844..7863)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(8528..9041)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8723..8744)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8935..8954
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      9152..9740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9641..9660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9894..9911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9926..10066
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9992..10011)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10152..10174
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(10168..10359)
                     /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 ggctagaaat aattttgttt
     5161 aactttaaga aggagatata catatgaaac aagatctgat gtcccctata ctaggttatt
     5221 ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt gaagaaaaat
     5281 atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat
     5341 tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa ttaacacagt
     5401 ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt tgtccaaaag
     5461 agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac ggtgtttcga
     5521 gaattgcata tagtaaagac tttgaaactc tcaaagttga ttttcttagc aagctacctg
     5581 aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat ggtgatcatg
     5641 taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac atggacccaa
     5701 tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa gctatcccac
     5761 aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag ggctggcaag
     5821 ccacgtttgg tggtggcgac catcctccaa ctagtggatc tggtggtggt ggcggttcca
     5881 tgagcgagaa tctttatttt caggtcgaca tggcgagtag cagcggctcc aaggctgaat
     5941 tcattgtcgg agggaaatat aaactggtac ggaagatcgg gtctggctcc ttcggggaca
     6001 tctatttggc gatcaacatc accaacggcg aggaagtggc agtgaagcta gaatctcaga
     6061 aggccaggca tccccagttg ctgtacgaga gcaagctcta taagattctt caaggtgggg
     6121 ttggcatccc ccacatacgg tggtatggtc aggaaaaaga ctacaatgta ctagtcatgg
     6181 atcttctggg acctagcctc gaagacctct tcaatttctg ttcaagaagg ttcacaatga
     6241 aaactgtact tatgttagct gaccagatga tcagtagaat tgaatatgtg catacaaaga
     6301 attttataca cagagacatt aaaccagata acttcctaat gggtattggg cgtcactgta
     6361 ataagttatt ccttattgat tttggtttgg ccaaaaagta cagagacaac aggacaaggc
     6421 aacacatacc atacagagaa gataaaaacc tcactggcac tgcccgatat gctagcatca
     6481 atgcacatct tggtattgag cagagtcgcc gagatgacat ggaatcatta ggatatgttt
     6541 tgatgtattt taatagaacc agcctgccat ggcaagggct aaaggctgca acaaagaaac
     6601 aaaaatatga aaagattagt gaaaagaaga tgtccacgcc tgttgaagtt ttatgtaagg
     6661 ggtttcctgc agaatttgcg atgtacttaa actattgtcg tgggctacgc tttgaggaag
     6721 ccccagatta catgtatctg aggcagctat tccgcattct tttcaggacc ctgaaccatc
     6781 aatatgacta cacatttgat tggacaatgt taaagcagaa agcagcacag caggcagcct
     6841 cttccagtgg gcagggtcag caggcccaaa cccccacagg caagcaaact gacaaatcca
     6901 agagtaacat gaaaggtttc tagctgcagt cgagaagctt ggcactggcc gtcgttttac
     6961 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     7021 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     7081 gcagcctgaa tggcgaatgg cagcttggct gttttggcgg atgagataag attttcagcc
     7141 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     7201 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     7261 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     7321 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     7381 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     7441 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     7501 acggatggcc tttttgcgtt tctacaaact ctttttgttt atttttctaa atacattcaa
     7561 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7621 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7681 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7741 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7801 gccccgaaga acgttctcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7861 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7921 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7981 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     8041 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     8101 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     8161 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     8221 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     8281 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     8341 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     8401 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     8461 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     8521 ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     8581 tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     8641 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagcc
     8701 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     8761 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     8821 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     8881 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     8941 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     9001 caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc taggtgaaga
     9061 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     9121 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     9181 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     9241 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     9301 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     9361 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     9421 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     9481 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     9541 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     9601 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     9661 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     9721 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     9781 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     9841 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     9901 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     9961 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
    10021 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
    10081 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
    10141 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
    10201 gaggcagctg cggtaaagct catcagcgtg gtcgtgcagc gattcacaga tgtctgcctg
    10261 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
    10321 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
    10381 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
    10441 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
    10501 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
    10561 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
    10621 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
    10681 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
    10741 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
    10801 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaaa tt
//
