LOCUS       sequence-258193-       16165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Dysferlin protein (Therapeutic gene) along with Follistatin 
            protein to cure LGMD2B along with luciferase enzyme for in vivo live
            imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16165)
  AUTHORS   Guha TK, Pichavant C, Calos MP
  TITLE     Plasmid-Mediated Gene Therapy in Mouse Models of Limb Girdle 
            Muscular Dystrophy.
  JOURNAL   Mol Ther Methods Clin Dev. 2019 Oct 14;15:294-304. doi: 
            10.1016/j.omtm.2019.10.002. eCollection 2019 Dec 13.
  PUBMED    31890729
REFERENCE   2  (bases 1 to 16165)
  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"; journalName: "Mol Ther 
            Methods Clin Dev."; date: "2019-10-14"; volume: "15:294-304. doi"; 
            pages: " 10.1016/j.omtm.2019.10.002. eCollection 2019 Dec 13"
COMMENT     Sequence Label: pKLD-CAG-DYSF-CFL
FEATURES             Location/Qualifiers
     source          1..16165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1407..2201
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1407..2201
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1461..1480)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2071..2090
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2526..3114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3015..3034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3178..3211
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3239..3308
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     enhancer        3564..3867
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3869..4143
                     /label=chicken beta-actin promoter
     intron          4144..5158
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5082..5104
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5166..5185
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5919..5930
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             11487..11543
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             11544..12251
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(11689..11707)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     11957..11976
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             12258..12314
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             13362..13418
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             13419..15071
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(13485..13503)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(15148..15167)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    15213..15268
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(15267..15286)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(15611..15630)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    15655..15702
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    15873..15922
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     primer_bind     complement(15935..15952)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    15941..16165
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gaattcaaac tagagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc
       61 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg
      121 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacaggatcg
      181 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg
      241 aaaaagcttt aggctgaaag agagatttag aatgacagaa tcatagaacg gcctgggttg
      301 caaaggagca cagtgctcat ccagatccaa ccccctgcta tgtgcagggt catcaaccag
      361 cagcccaggc tgcccagagc cacatccagc ctggccttga atgcctgcag ggatggggca
      421 tccacagcct ccttgggcaa cctgttcagt gcgtcaccac cctctggggg aaaaactgcc
      481 tcctcatatc caacccaaac ctcccctgtc tcagtgtaaa gccattcccc cttgtcctat
      541 caagggggag tttgctgtga cattgttggt ctggggtgac acatgtttgc caattcagtg
      601 catcacggag aggcagatct tggggataag gaagtgcagg acagcatgga cgtgggacat
      661 gcaggtgttg agggctctgg gacactctcc aagtcacagc gttcagaaca gccttaaaga
      721 taagaagata ggatagaagg acaaagagca agttaaaacc cagcatggag aggagcacaa
      781 aaaggccaca gacactgctg gtccctgtgt ctgagcctgc atgtttgatg gtgtctggat
      841 gcaagcagaa ggggtggaag agcttgcctg gagagataca gctgggtcag taggactggg
      901 acaggcagct ggagaattgc catgtagatg ttcatacaat cgtcaaatca tgaaggctgg
      961 aaaagccctc caagatcccc aagaccaacc ccaacccacc caccgtgccc actggccatg
     1021 tccctcagtg ccacatcccc acagttcttc atcacctcca gggacggtga cccccccacc
     1081 tccgtgggca gctgtgccac tgcagcaccg ctctttggag aaggtaaatc ttgctaaatc
     1141 cagcccgacc ctcccctggc acaacgtaag gccattatct ctcatccaac tccaggacgg
     1201 agtcagtgag gatggggctc tagtttgcgc gcctcttcta ctgggcggtt ttatggacag
     1261 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag
     1321 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agctctgatc
     1381 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     1441 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     1501 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     1561 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     1621 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     1681 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     1741 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     1801 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     1861 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     1921 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     1981 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     2041 tgggggtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     2101 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     2161 agcgcatcgc cttctatcgc cttcttgacg agttcttctg aattattaac gcttacaatt
     2221 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atcaggtggc
     2281 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2341 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatagca cgtgctaaaa
     2401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2641 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcata
     3181 acttcgtata atgtatgcta tacgaagtta tcttaaggtc gacgatgtag gtcacggtct
     3241 cgaagccgcg gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg tactccacct
     3301 cacccatctg gtccatcatg atgaacgggt cgaggtggcg gtagttgatc ccggcgaacg
     3361 cgcggcgcac cgggaagccc tcgccctcga aaccgctggg cgcggtggtc acggtgagca
     3421 cgggacgtgc gacggcgtcg gcgggtgcgg atacgcgggg cagcgtcagc gggttctcga
     3481 cggtcacggc gggcatgtcg acctcgagat taatagtaat caattacggg gtcattagtt
     3541 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     3601 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     3661 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     3721 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     3781 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3841 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
     3901 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
     3961 gcagcgatgg gggcgggggg gggggggggc gcgccaggcg gggcggggcg gggcgagggg
     4021 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     4081 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     4141 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     4201 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     4261 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
     4321 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
     4381 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
     4441 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     4501 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     4561 gtgtgcgtgg gggggtgagc agggggtggg cgcgtcggtc gggctgcaac cccccctgca
     4621 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     4681 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     4741 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     4801 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     4861 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     4921 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     4981 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     5041 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     5101 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     5161 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccgt
     5221 ttaaacgatc ggccgccacc atgctgaggg tcttcatcct ctatgccgag aacgtccaca
     5281 cacccgacac cgacatcagc gatgcctact gctccgcggt gtttgcaggg gtgaagaaga
     5341 gaaccaaagt catcaagaac agcgtgaacc ctgtatggaa tgagggattt gaatgggacc
     5401 tcaagggcat ccccctggac cagggctctg agcttcatgt ggtggtcaaa gaccatgaga
     5461 cgatggggag gaacaggttc ctgggggaag ccaaggtccc actccgagag gtcctcgcca
     5521 cccctagtct gtccgccagc ttcaatgccc ccctgctgga caccaagaag cagcccacag
     5581 gggcctcgct ggtcctgcag gtgtcctaca caccgctgcc tggagctgtg cccctgttcc
     5641 cgccccctac tcctctggag ccctccccga ctctgcctga cctggatgta gtggcagaca
     5701 caggaggaga ggaagacaca gaggaccagg gactcactgg agatgaggcg gagccattcc
     5761 tggatcaaag cggaggcccg ggggctccca ccaccccaag gaaactacct tcacgtcctc
     5821 cgccccacta ccccgggatc aaaagaaagc gaagtgcgcc tacatctaga aagctgctgt
     5881 cagacaaacc gcaggatttc cagatcaggg tccaggtgat cgaggggcgc cagctgccgg
     5941 gggtgaacat caagcctgtg gtcaaggtta ccgctgcagg gcagaccaag cggacgcgga
     6001 tccacaaggg aaacagccca ctcttcaatg agactctttt cttcaacttg tttgactctc
     6061 ctggggagct gtttgatgag cccatcttta tcacggtggt agactctcgt tctctcagga
     6121 cagatgctct cctcggggag ttccggatgg acgtgggcac catttacaga gagccccggc
     6181 acgcctatct caggaagtgg ctgctgctct cagaccctga tgacttctct gctggggcca
     6241 gaggctacct gaaaacaagc ctttgtgtgc tggggcctgg ggacgaagcg cctctggaga
     6301 gaaaagaccc ctctgaagac aaggaggaca ttgaaagcaa cctgctccgg cccacaggcg
     6361 tagccctgcg aggagcccac ttctgcctga aggtcttccg ggccgaggac ttgccgcaga
     6421 tggacgatgc cgtgatggac aacgtgaaac agatctttgg cttcgagagt aacaagaaga
     6481 acttggtgga cccctttgtg gaggtcagct ttgcggggaa aatgctgtgc agcaagatct
     6541 tggagaagac ggccaaccct cagtggaacc agaacatcac actgcctgcc atgtttccct
     6601 ccatgtgcga aaaaatgagg attcgtatca tagactggga ccgcctgact cacaatgaca
     6661 tcgtggctac cacctacctg agtatgtcga aaatctctgc ccctggagga gaaatagaag
     6721 aggagcctgc aggtgctgtc aagccttcga aagcctcaga cttggatgac tacctgggct
     6781 tcctccccac ttttgggccc tgctacatca acctctatgg cagtcccaga gagttcacag
     6841 gcttcccaga cccctacaca gagctcaaca caggcaaggg ggaaggtgtg gcttatcgtg
     6901 gccggcttct gctctccctg gagaccaagc tggtggagca cagtgaacag aaggtggagg
     6961 accttcctgc ggatgacatc ctccgggtgg agaagtacct taggaggcgc aagtactccc
     7021 tgtttgcggc cttctactca gccaccatgc tgcaggatgt ggatgatgcc atccagtttg
     7081 aggtcagcat cgggaactac gggaacaagt tcgacatgac ctgcctgccg ctggcctcca
     7141 ccactcagta cagccgtgca gtctttgacg ggtgccacta ctactaccta ccctggggta
     7201 acgtgaaacc tgtggtggtg ctgtcatcct actgggagga catcagccat agaatcgaga
     7261 ctcagaacca gctgcttggg attgctgacc ggctggaagc tggcctggag caggtccacc
     7321 tggccctgaa ggcgcagtgc tccacggagg acgtggactc gctggtggct cagctgacgg
     7381 atgagctcat cgcaggctgc agccagcctc tgggtgacat ccatgagaca ccctctgcca
     7441 cccacctgga ccagtacctg taccagctgc gcacccatca cctgagccaa atcactgagg
     7501 ctgccctggc cctgaagctc ggccacagtg agctccctgc agctctggag caggcggagg
     7561 actggctcct gcgtctgcgt gccctggcag aggagcccca gaacagcctg ccggacatcg
     7621 tcatctggat gctgcaggga gacaagcgtg tggcatacca gcgggtgccc gcccaccaag
     7681 tcctcttctc ccggcggggt gccaactact gtggcaagaa ttgtgggaag ctacagacaa
     7741 tctttctgaa atatccgatg gagaaggtgc ctggcgcccg gatgccagtg cagatacggg
     7801 tcaagctgtg gtttgggctc tctgtggatg agaaggagtt caaccagttt gctgagggga
     7861 agctgtctgt ctttgctgaa acctatgaga acgagactaa gttggccctt gttgggaact
     7921 ggggcacaac gggcctcacc taccccaagt tttctgacgt cacgggcaag atcaagctac
     7981 ccaaggacag cttccgcccc tcggccggct ggacctgggc tggagattgg ttcgtgtgtc
     8041 cggagaagac tctgctccat gacatggacg ccggtcacct gagcttcgtg gaagaggtgt
     8101 ttgagaacca gacccggctt cccggaggcc agtggatcta catgagtgac aactacaccg
     8161 atgtgaacgg ggagaaggtg cttcccaagg atgacattga gtgcccactg ggctggaagt
     8221 gggaagatga ggaatggtcc acagacctca accgggctgt cgatgagcaa ggctgggagt
     8281 atagcatcac catccccccg gagcggaagc cgaagcactg ggtccctgct gagaagatgt
     8341 actacacaca ccgacggcgg cgctgggtgc gcctgcgcag gagggatctc agccaaatgg
     8401 aagcactgaa aaggcacagg caggcggagg cggagggcga gggctgggag tacgcctctc
     8461 tttttggctg gaagttccac ctcgagtacc gcaagacaga tgccttccgc cgccgccgct
     8521 ggcgccgtcg catggagcca ctggagaaga cggggcctgc agctgtgttt gcccttgagg
     8581 gggccctggg cggcgtgatg gatgacaaga gtgaagattc catgtccgtc tccaccttga
     8641 gcttcggtgt gaacagaccc acgatttcct gcatattcga ctatgggaac cgctaccatc
     8701 tacgctgcta catgtaccag gcccgggacc tggctgcgat ggacaaggac tctttttctg
     8761 atccctatgc catcgtctcc ttcctgcacc agagccagaa gacggtggtg gtgaagaaca
     8821 cccttaaccc cacctgggac cagacgctca tcttctacga gatcgagatc tttggcgagc
     8881 cggccacagt tgctgagcaa ccgcccagca ttgtggtgga gctgtacgac catgacactt
     8941 atggtgcaga cgagtttatg ggtcgctgca tctgtcaacc gagtctggaa cggatgccac
     9001 ggctggcctg gttcccactg acgaggggca gccagccgtc gggggagctg ctggcctctt
     9061 ttgagctcat ccagagagag aagccggcca tccaccatat tcctggtttt gaggtgcagg
     9121 agacatcaag gatcctggat gagtctgagg acacagacct gccctaccca ccaccccaga
     9181 gggaggccaa catctacatg gttcctcaga acatcaagcc agcgctccag cgtaccgcca
     9241 tcgagatcct ggcatggggc ctgcggaaca tgaagagtta ccagctggcc aacatctcct
     9301 cccccagcct cgtggtagag tgtgggggcc agacggtgca gtcctgtgtc atcaggaacc
     9361 tccggaagaa ccccaacttt gacatctgca ccctcttcat ggaagtgatg ctgcccaggg
     9421 aggagctcta ctgccccccc atcaccgtca aggtcatcga taaccgccag tttggccgcc
     9481 ggcctgtggt gggccagtgt accatccgct ccctggagag cttcctgtgt gacccctact
     9541 cggcggagag tccatcccca cagggtggcc cagacgatgt gagcctactc agtcctgggg
     9601 aagacgtgct catcgacatt gatgacaagg agcccctcat ccccatccag gaggaagagt
     9661 tcatcgattg gtggagcaaa ttctttgcct ccatagggga gagggaaaag tgcggctcct
     9721 acctggagaa ggattttgac accctgaagg tctatgacac acagctggag aatgtggagg
     9781 cctttgaggg cctgtctgac ttttgtaaca ccttcaagct gtaccggggc aagacgcagg
     9841 aggagacaga agatccatct gtgattggtg aatttaaggg cctcttcaaa atttatcccc
     9901 tcccagaaga cccagccatc cccatgcccc caagacagtt ccaccagctg gccgcccagg
     9961 gaccccagga gtgcttggtc cgtatctaca ttgtccgagc atttggcctg cagcccaagg
    10021 accccaatgg aaagtgtgat ccttacatca agatctccat agggaagaaa tcagtgagtg
    10081 accaggataa ctacatcccc tgcacgctgg agcccgtatt tggaaagatg ttcgagctga
    10141 cctgcactct gcctctggag aaggacctaa agatcactct ctatgactat gacctcctct
    10201 ccaaggacga aaagatcggt gagacggtcg tcgacctgga gaacaggctg ctgtccaagt
    10261 ttggggctcg ctgtggactc ccacagacct actgtgtctc tggaccgaac cagtggcggg
    10321 accagctccg cccctcccag ctcctccacc tcttctgcca gcagcataga gtcaaggcac
    10381 ctgtgtaccg gacagaccgt gtaatgtttc aggataaaga atattccatt gaagagatag
    10441 aggctggcag gatcccaaac ccacacctgg gcccagtgga ggagcgtctg gctctgcatg
    10501 tgcttcagca gcagggcctg gtcccggagc acgtggagtc acggcccctc tacagccccc
    10561 tgcagccaga catcgagcag gggaagctgc agatgtgggt cgacctattt ccgaaggccc
    10621 tggggcggcc tggacctccc ttcaacatca ccccacggag agccagaagg tttttcctgc
    10681 gttgtattat ctggaatacc agagatgtga tcctggatga cctgagcctc acgggggaga
    10741 agatgagcga catttatgtg aaaggttgga tgattggctt tgaagaacac aagcaaaaga
    10801 cagacgtgca ttatcgttcc ctgggaggtg aaggcaactt caactggagg ttcattttcc
    10861 ccttcgacta cctgccagct gagcaagtct gtaccattgc caagaaggat gccttctgga
    10921 ggctggacaa gactgagagc aaaatcccag cacgagtggt gttccagatc tgggacaatg
    10981 acaagttctc ctttgatgat tttctgggct ccctgcagct cgatctcaac cgcatgccca
    11041 agccagccaa gacagccaag aagtgctcct tggaccagct ggatgatgct ttccacccag
    11101 aatggtttgt gtcccttttt gagcagaaaa cagtgaaggg ctggtggccc tgtgtagcag
    11161 aagagggtga gaagaaaata ctggcgggca agctggaaat gaccttggag attgtagcag
    11221 agagtgagca tgaggagcgg cctgctggcc agggccggga tgagcccaac atgaacccta
    11281 agcttgagga cccaaggcgc cccgacacct ccttcctgtg gtttacctcc ccatacaaga
    11341 ccatgaagtt catcctgtgg cggcgtttcc ggtgggccat catcctcttc atcatcctct
    11401 tcatcctgct gctgttcctg gccatcttca tctacgcctt cccgaactat gctgccatga
    11461 agctggtgaa gcccttcagc acgcgtgcca cgaacttctc tctgttaaag caagcaggag
    11521 acgtggaaga aaaccccggt cctatggtga gcaagggcga ggaggataac atggccatca
    11581 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
    11641 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
    11701 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
    11761 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
    11821 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
    11881 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
    11941 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
    12001 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
    12061 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
    12121 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
    12181 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
    12241 agctgtacaa gaccggtgcc acgaacttct ctctgttaaa gcaagcagga gacgtggaag
    12301 aaaaccccgg tcctatggtc cgcgcgaggc accagccggg tgggctttgc ctcctgctgc
    12361 tgctgctctg ccagttcatg gaggaccgca gtgcccaggc tgggaactgc tggctccgtc
    12421 aagcgaagaa cggccgctgc caggtcctgt acaagaccga actgagcaag gaggagtgct
    12481 gcagcaccgg ccggctgagc acctcgtgga ccgaggagga cgtgaatgac aacacactct
    12541 tcaagtggat gattttcaac gggggcgccc ccaactgcat cccctgtaaa gaaacgtgtg
    12601 agaacgtgga ctgtggacct gggaaaaaat gccgaatgaa caagaagaac aaaccccgct
    12661 gcgtctgcgc cccggattgt tccaacatca cctggaaggg tccagtctgc gggctggatg
    12721 ggaaaaccta ccgcaatgaa tgtgcactcc taaaggcaag atgtaaagag cagccagaac
    12781 tggaagtcca gtaccaaggc agatgtaaaa agacttgtcg ggatgttttc tgtccaggca
    12841 gctccacatg tgtggtggac cataccaata atgcctactg tgtgacctgt aatcggattt
    12901 gcccagagcc tgcttcctct gagcaatatc tctgtgggaa tgatggagtc acctactcca
    12961 gtgcctgcca cctaagaaag gctacctgcc tgctgggcag atctattgga ttagcctatg
    13021 agggaaagtg tatcaaagca aagtcctgtg aagatatcca gtgcactggt gggaaaaaat
    13081 gtttatggga tttcaaggtt gggagaggcc ggtgttccct ctgtgatgag ctgtgccctg
    13141 acagtaagtc ggatgagcct gtctgtgcca gtgacaatgc cacttatgcc agcgagtgtg
    13201 ccatgaagga agctgcctgc tcctcaggtg tgctactgga agtaaagcac tccggatctt
    13261 gcaactccat ttcggaagac accgaggaag aggaggaaga tgaagaccag gactacagct
    13321 ttcctatatc ttctattcta gagtggaccg gtggatccgg tgccacgaac ttctctctgt
    13381 taaagcaagc aggagacgtg gaagaaaacc ccggtcctat ggaagacgcc aaaaacataa
    13441 agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata
    13501 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
    13561 aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga
    13621 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
    13681 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
    13741 tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg
    13801 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa
    13861 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
    13921 tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata
    13981 gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg
    14041 tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg
    14101 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
    14161 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
    14221 ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg
    14281 tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
    14341 atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg
    14401 ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag
    14461 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
    14521 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag
    14581 gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga
    14641 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
    14701 aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc
    14761 aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag
    14821 gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
    14881 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
    14941 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
    15001 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga
    15061 tcgccgtgta agcggccgcg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
    15121 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
    15181 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
    15241 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
    15301 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
    15361 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
    15421 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
    15481 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
    15541 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctctctagag gatcctaggg
    15601 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
    15661 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
    15721 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
    15781 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
    15841 acgtcggggc ggcaggccct gccatagcct cgtcggccgg cttgtcgacg acggcggtct
    15901 ccgtcgtcag gatcatccgg gccacacgcc tagccctcga ctgtgccttc tagttgccag
    15961 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
    16021 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
    16081 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
    16141 gctggggatg cggtgggctc tatgg
//
