LOCUS       sequence-308806-       10851 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 10851)
  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 10851)
  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..10851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..78
                     /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             79..1161
                     /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(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1404..1409,1410..1425,1426..1432)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1440..1456
                     /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     1445..1467
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1526..2626
                     /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="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      2483..2492
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2600..2623
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2675..2686
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2729..2749
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2753..5095
                     /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             2753..5095
                     /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             5105..5122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5152..5174
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5197..5847
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
     primer_bind     5808..5830
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     primer_bind     complement(6942..6959)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6942..6958)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6951..6973)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7137..7154)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      7307..7393
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7485..7512
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        7531..7622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7623..7691,7692..8483)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7841..7860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(8525..9038)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8720..8741)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8932..8951
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      9149..9737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9638..9657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9891..9908
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9923..10063
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9989..10008)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10149..10171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(10165..10356)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     1201 acaattctca tgtttgacag cttatcatcg actgcacggt gcaccaatgc ttctggcgtc
     1261 aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg
     1321 tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca taacggttct
     1381 ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa tgtgtggaat
     1441 tgtgagcgga taacaatttc acacaggaaa cagccagtcc gtttaggtgt tttcacgagc
     1501 acttcaccaa caaggaccat agattatgaa aactgaagaa ggtaaactgg taatctggat
     1561 taacggcgat aaaggctata acggtctcgc tgaagtcggt aagaaattcg agaaagatac
     1621 cggaattaaa gtcaccgttg agcatccgga taaactggaa gagaaattcc cacaggttgc
     1681 ggcaactggc gatggccctg acattatctt ctgggcacac gaccgctttg gtggctacgc
     1741 tcaatctggc ctgttggctg aaatcacccc ggacaaagcg ttccaggaca agctgtatcc
     1801 gtttacctgg gatgccgtac gttacaacgg caagctgatt gcttacccga tcgctgttga
     1861 agcgttatcg ctgatttata acaaagatct gctgccgaac ccgccaaaaa cctgggaaga
     1921 gatcccggcg ctggataaag aactgaaagc gaaaggtaag agcgcgctga tgttcaacct
     1981 gcaagaaccg tacttcacct ggccgctgat tgctgctgac gggggttatg cgttcaagta
     2041 tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat aacgctggcg cgaaagcggg
     2101 tctgaccttc ctggttgacc tgattaaaaa caaacacatg aatgcagaca ccgattactc
     2161 catcgcagaa gctgccttta ataaaggcga aacagcgatg accatcaacg gcccgtgggc
     2221 atggtccaac atcgacacca gcaaagtgaa ttatggtgta acggtactgc cgaccttcaa
     2281 gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg ccgccagtcc
     2341 gaacaaagag ctggcaaaag agttcctcga aaactatctg ctgactgatg aaggtctgga
     2401 agcggttaat aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg aggaagagtt
     2461 ggcgaaagat ccacgtattg ccgccaccat ggaaaacgcc cagaaaggtg aaatcatgcc
     2521 gaacatcccg cagatgtccg ctttctggta tgccgtgcgt actgcggtga tcaacgccgc
     2581 cagcggtcgt cagactgtcg atgaagccct gaaagacgcg cagactaatt cgagctcgaa
     2641 caacaacaac aataacaata acaacaacct cgggatcgag ggaaggattt caactagtgg
     2701 atctggtggt ggtggcggtt ccatgagcga gaatctttat tttcagggat ccatggctac
     2761 tcaagctgat ttgatggagt tggacatggc catggaacca gacagaaaag cggctgttag
     2821 tcactggcag caacagtctt acctggactc tggaatccat tctggtgcca ctaccacagc
     2881 tccttctctg agtggtaaag gcaatcctga ggaagaggat gtggatacct cccaagtcct
     2941 gtatgagtgg gaacagggat tttctcagtc cttcactcaa gaacaagtag ctgatattga
     3001 tggacagtat gcaatgactc gagctcagag ggtacgagct gctatgttcc ctgagacatt
     3061 agatgagggc atgcagatcc catctacaca gtttgatgct gctcatccca ctaatgtcca
     3121 gcgtttggct gaaccatcac agatgctgaa acatgcagtt gtaaacttga ttaactatca
     3181 agatgatgca gaacttgcca cacgtgcaat ccctgaactg acaaaactgc taaatgacga
     3241 ggaccaggtg gtggttaata aggctgcagt tatggtccat cagctttcta aaaaggaagc
     3301 ttccagacac gctatcatgc gttctcctca gatggtgtct gctattgtac gtaccatgca
     3361 gaatacaaat gatgtagaaa cagctcgttg taccgctggg accttgcata acctttccca
     3421 tcatcgtgag ggcttactgg ccatctttaa gtctggaggc attcctgccc tggtgaaaat
     3481 gcttggttca ccagtggatt ctgtgttgtt ttatgccatt acaactctcc acaacctttt
     3541 attacatcaa gaaggagcta aaatggcagt gcgtttagct ggtgggctgc agaaaatggt
     3601 tgccttgctc aacaaaacaa atgttaaatt cttggctatt acgacagact gccttcaaat
     3661 tttagcttat ggcaaccaag aaagcaagct catcatactg gctagtggtg gaccccaagc
     3721 tttagtaaat ataatgagga cctatactta cgaaaaacta ctgtggacca caagcagagt
     3781 gctgaaggtg ctatctgtct gctctagtaa taagccggct attgtagaag ctggtggaat
     3841 gcaagcttta ggacttcacc tgacagatcc aagtcaacgt cttgttcaga actgtctttg
     3901 gactctcagg aatctttcag atgctgcaac taaacaggaa gggatggaag gtctccttgg
     3961 gactcttgtt cagcttctgg gttcagatga tataaatgtg gtcacctgtg cagctggaat
     4021 tctttctaac ctcacttgca ataattataa gaacaagatg atggtctgcc aagtgggtgg
     4081 tatagaggct cttgtgcgta ctgtccttcg ggctggtgac agggaagaca tcactgagcc
     4141 tgccatctgt gctcttcgtc atctgaccag ccgacaccaa gaagcagaga tggcccagaa
     4201 tgcagttcgc cttcactatg gactaccagt tgtggttaag ctcttacacc caccatccca
     4261 ctggcctctg ataaaggcta ctgttggatt gattcgaaat cttgcccttt gtcccgcaaa
     4321 tcatgcacct ttgcgtgagc agggtgccat tccacgacta gttcagttgc ttgttcgtgc
     4381 acatcaggat acccagcgcc gtacgtccat gggtgggaca cagcagcaat ttgtggaggg
     4441 ggtccgcatg gaagaaatag ttgaaggttg taccggagcc cttcacatcc tagctcggga
     4501 tgttcacaac cgaattgtta tcagaggact aaataccatt ccattgtttg tgcagctgct
     4561 ttattctccc attgaaaaca tccaaagagt agctgcaggg gtcctctgtg aacttgctca
     4621 ggacaaggaa gctgcagaag ctattgaagc tgagggagcc acagctcctc tgacagagtt
     4681 acttcactct aggaatgaag gtgtggcgac atatgcagct gctgttttgt tccgaatgtc
     4741 tgaggacaag ccacaagatt acaagaaacg gctttcagtt gagctgacca gctctctctt
     4801 cagaacagag ccaatggctt ggaatgagac tgctgatctt ggacttgata ttggtgccca
     4861 gggagaaccc cttggatatc gccaggatga tcctagctat cgttcttttc actctggtgg
     4921 atatggccag gatgccttgg gtatggaccc catgatggaa catgagatgg gtggccacca
     4981 ccctggtgct gactatccag ttgatgggct gccagatctg gggcatgccc aggacctcat
     5041 ggatgggctg cctccaggtg acagcaatca gctggcctgg tttgatactg acctgagcgg
     5101 ccgccaccat caccaccacc actaggggcc gcactcgagg ctagaaataa ttttgtttaa
     5161 ctttaagaag gagatataca tatgaaacaa gatctgatgt cccctatact aggttattgg
     5221 aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat
     5281 gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg
     5341 ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct
     5401 atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag
     5461 cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga
     5521 attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa
     5581 atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta
     5641 acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg
     5701 tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa
     5761 attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc
     5821 acgtttggtg gtggcgacca tcctccaact agtggatctg gtggtggtgg cggttccatg
     5881 agcgagaatc tttattttca ggtcgacatg gcgagtagca gcggctccaa ggctgaattc
     5941 attgtcggag ggaaatataa actggtacgg aagatcgggt ctggctcctt cggggacatc
     6001 tatttggcga tcaacatcac caacggcgag gaagtggcag tgaagctaga atctcagaag
     6061 gccaggcatc cccagttgct gtacgagagc aagctctata agattcttca aggtggggtt
     6121 ggcatccccc acatacggtg gtatggtcag gaaaaagact acaatgtact agtcatggat
     6181 cttctgggac ctagcctcga agacctcttc aatttctgtt caagaaggtt cacaatgaaa
     6241 actgtactta tgttagctga ccagatgatc agtagaattg aatatgtgca tacaaagaat
     6301 tttatacaca gagacattaa accagataac ttcctaatgg gtattgggcg tcactgtaat
     6361 aagttattcc ttattgattt tggtttggcc aaaaagtaca gagacaacag gacaaggcaa
     6421 cacataccat acagagaaga taaaaacctc actggcactg cccgatatgc tagcatcaat
     6481 gcacatcttg gtattgagca gagtcgccga gatgacatgg aatcattagg atatgttttg
     6541 atgtatttta atagaaccag cctgccatgg caagggctaa aggctgcaac aaagaaacaa
     6601 aaatatgaaa agattagtga aaagaagatg tccacgcctg ttgaagtttt atgtaagggg
     6661 tttcctgcag aatttgcgat gtacttaaac tattgtcgtg ggctacgctt tgaggaagcc
     6721 ccagattaca tgtatctgag gcagctattc cgcattcttt tcaggaccct gaaccatcaa
     6781 tatgactaca catttgattg gacaatgtta aagcagaaag cagcacagca ggcagcctct
     6841 tccagtgggc agggtcagca ggcccaaacc cccacaggca agcaaactga caaatccaag
     6901 agtaacatga aaggtttcta gctgcagtcg agaagcttgg cactggccgt cgttttacaa
     6961 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     7021 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     7081 agcctgaatg gcgaatggca gcttggctgt tttggcggat gagataagat tttcagcctg
     7141 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     7201 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     7261 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     7321 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     7381 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     7441 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     7501 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
     7561 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7621 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     7681 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     7741 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     7801 ccgaagaacg tttcccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7861 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7921 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7981 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     8041 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     8101 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     8161 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     8221 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     8281 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     8341 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     8401 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     8461 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     8521 atttaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat aagcaaatat
     8581 ttaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     8641 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
     8701 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     8761 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     8821 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     8881 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     8941 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     9001 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtaa aaggatctag gtgaagatcc
     9061 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     9121 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     9181 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     9241 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     9301 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     9361 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     9421 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     9481 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     9541 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     9601 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     9661 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     9721 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     9781 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     9841 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     9901 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     9961 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
    10021 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
    10081 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
    10141 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
    10201 gcagctgcgg taaagctcat cagcgtggtc gtgcagcgat tcacagatgt ctgcctgttc
    10261 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
    10321 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aatggggatt
    10381 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
    10441 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
    10501 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
    10561 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
    10621 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
    10681 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
    10741 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
    10801 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgaaattc c
//
