LOCUS       sequence-371826-       13278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9-KRAB-MeCP2 expression 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13278)
  AUTHORS   Nghi Dang , Alissa Lance-Byrne, Angela Tung, Xiaoge Guo, Ryan J 
            Cecchi, Joanna Buchthal, Alejandro Chavez, Nan Cher Yeo
  TITLE     Generation and application of a versatile CRISPR toolkit for 
            mammalian cell engineering
  JOURNAL   Synthetic Biology, Volume 6, Issue 1, 2021
REFERENCE   2  (bases 1 to 13278)
  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: "Synthetic 
            Biology, Volume 6, Issue 1, 2021"
COMMENT     Sequence Label: NAX_CAGGS_CAS9_m4_KRAB-MeCP2_KanR neo
FEATURES             Location/Qualifiers
     source          1..13278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1..1017
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     941..963
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1025..1044
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1114..1123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1120..1146,1147..1149,1150..3633,3634..3639,3640..3705,
                     3706..3708,3709..5223)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5236..5256
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5317..5511
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             5545..5565
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5575..6432
                     /codon_start=1
                     /product="transcriptional repression domain from rat 
                     methyl-CpG-binding protein 2 (Nan et al., 1997)"
                     /label=MeCP2
                     /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP
                     GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE
                     TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH
                     HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP
                     KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV
                     TERVS"
     primer_bind     complement(6487..6506)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6531..6579
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6810..7021
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             7037..7831
                     /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             7037..7831
                     /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(7091..7110)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7701..7720
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7835..7956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7872..7891)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7926..7945
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8719..8753
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9244..9260)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9244..9260)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9257..9279)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9268..9284
                     /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)."
     promoter        complement(join(9292..9298,9299..9316,9317..9322))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9337..9358
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9475..9492)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9646..10234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9726..9745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10405..11196,11197..11265))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     11028..11047
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11266..11370)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11396..11851)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11533..11554)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11745..11764
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11978..12000
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11992..12009
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11993..12009
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   12139..12201
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        12622..13001
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
       61 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      121 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      181 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      241 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      301 tgcgggcgcg gcgcggggct ttttgcgctc cgcagtgtgc gcgaggggag cgcggccggg
      361 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
      421 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
      481 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
      541 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg kgggggtgcc gggcggggcg
      601 gggccgccyc gggccggggg gggckcgggg grggggcgcg gcggcccccg gagcgccggc
      661 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
      721 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
      781 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
      841 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
      901 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
      961 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1021 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatc tagataccct
     1081 cgtaaaggtc tagagctagc ggatcgaacc cttgccacca tggacaagaa gtactccatt
     1141 gggctcgcta tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg
     1201 ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc
     1261 attggcgccc tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca
     1321 gcacggcgca gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt
     1381 aatgagatgg ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg
     1441 gaggaggata aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg
     1501 taccatgaaa agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat
     1561 aaggctgact tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac
     1621 ttcctcatcg agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa
     1681 ctggttcaga cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac
     1741 gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca
     1801 cagctccctg gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg
     1861 ctgaccccca actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc
     1921 aaagacacct acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca
     1981 gacctttttt tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga
     2041 gtgaacacgg agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag
     2101 caccaccaag acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac
     2161 aaggaaattt tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca
     2221 agccaggagg aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag
     2281 gagctgctgg taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat
     2341 ggaagcatcc cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag
     2401 gatttctacc cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg
     2461 ataccctact atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc
     2521 aaatcagaag agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct
     2581 gcccagtcct tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg
     2641 cttcctaaac actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc
     2701 aaatacgtca cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct
     2761 atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac
     2821 tatttcaaaa agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc
     2881 aacgcatccc tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg
     2941 gacaatgagg agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa
     3001 gatagggaga tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc
     3061 atgaaacagc tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc
     3121 aatgggatcc gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga
     3181 tttgccaacc ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac
     3241 atccagaaag cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt
     3301 gcaggtagcc cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc
     3361 gtcaaagtaa tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac
     3421 caaactaccc agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt
     3481 ataaaagaac tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag
     3541 aatgagaagc tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa
     3601 ctggacatca atcggctctc cgactacgac gtggctgcta tcgtgcccca gtcttttctc
     3661 aaagatgatt ctattgataa taaagtgttg acaagatccg ataaagctag agggaagagt
     3721 gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg
     3781 aacgccaaac tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc
     3841 ctgtctgagt tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc
     3901 accaagcacg tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac
     3961 aaactgattc gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga
     4021 aaggactttc agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc
     4081 tacctgaatg cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa
     4141 tttgtttacg gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag
     4201 gaaataggca aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag
     4261 accgagatta cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga
     4321 gaaacaggag aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg
     4381 tccatgccgc aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag
     4441 gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac
     4501 cccaagaaat acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc
     4561 aaagtggaga aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca
     4621 atcatggagc gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat
     4681 aaagaggtca aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa
     4741 aacggccgga aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca
     4801 ctgccctcta aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg
     4861 tctcccgaag ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat
     4921 gagatcatcg agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc
     4981 gataaggtgc tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa
     5041 aacattatcc acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc
     5101 gacaccacca tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg
     5161 attcatcagt caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga
     5221 gacagcaggg ctgaccccaa gaagaagagg aaggtgagtg gtggaggaag tggcgggtca
     5281 gggtcgatgg acgcgaaatc acttacggca tggtcgagaa cactggttac gttcaaggac
     5341 gtgtttgtgg actttacacg tgaggagtgg aaattgctgg atactgcgca acaaattgtg
     5401 tatcgaaatg tcatgcttga gaattacaag aacctcgtca gtctcggata ccagttgacg
     5461 aaaccggatg tgatccttag gctcgaaaag ggggaagaac cttggctggt atcgggaggt
     5521 ggttcgggtg gctctggatc aagcccaaag aagaaacgga aggtggaagc ctcagtgcag
     5581 gtgaaaaggg tgctggaaaa atcccccggc aaactcctcg tgaagatgcc cttccaggct
     5641 tcccctggcg gaaaaggtga agggggtggc gcaaccacat ctgcccaggt catggtcatc
     5701 aagcgacctg gaaggaaaag aaaggccgag gctgaccctc aggccattcc aaagaaacgg
     5761 ggacgcaagc cagggtccgt ggtcgcagct gcagcagctg aggctaagaa aaaggcagtg
     5821 aaggaaagct ccatccgcag tgtgcaggag actgtcctgc ccatcaagaa gaggaagact
     5881 agggagaccg tgtccatcga ggtcaaagaa gtggtcaagc ccctgctcgt gtccaccctg
     5941 ggcgaaaaat ctggaaaggg gctcaaaaca tgcaagtcac ctggacggaa aagcaaggag
     6001 tctagtccaa aggggcgctc aagctccgct tctagtcccc ctaaaaagga acaccatcac
     6061 catcaccatc acgccgagtc tcctaaggct cctatgccac tgctcccacc acctccacca
     6121 cctgagccac agtcaagcga agaccccatc agcccacccg agcctcagga tctgtcctct
     6181 agtatttgca aagaggaaaa gatgcccaga gcaggcagcc tggagagtga tggctgtcca
     6241 aaagaacccg ccaagaccca gcctatggtg gcagccgctg caactaccac cacaaccaca
     6301 actaccacag tggccgaaaa atacaagcat cgcggcgagg gcgaacgaaa ggacattgtg
     6361 tcaagctcca tgcccagacc taaccgggag gaaccagtcg atagtaggac acccgtgact
     6421 gagagagtct catagaagaa ttcgatccct accggttagt aatgagttta aacgggggag
     6481 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     6541 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6601 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6661 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6721 tcggggcggc aggccctgcc atagcagatc tgcgaaatcg gatccgcgcc gcactgattc
     6781 tagctgcaga ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     6841 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     6901 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     6961 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     7021 ggatcctcga gccaccatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     7081 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     7141 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     7201 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     7261 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     7321 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     7381 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     7441 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     7501 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     7561 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     7621 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     7681 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     7741 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     7801 cttctatcgc cttcttgacg agttcttcta agttaacttg tttattgcag cttataatgg
     7861 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     7921 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggaatt gactcaaatg
     7981 atgtcaatta gtctatcaga agctcatctg gtctcccttc cgggggacaa gacatccctg
     8041 tttaatattt aaacagcagt gttcccaaac tgggttctta tatcccttgc tctggtcaac
     8101 caggttgcag ggtttcctgt cctcacagga acgaagtccc taaagaaaca gtggcagcca
     8161 ggtttagccc cggaattgac tggattcctt ttttagggcc cattggtatg gctttttccc
     8221 cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga
     8281 tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg
     8341 ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag
     8401 ggacgtaatt acatccctgg gggctttggg ggggggctgt ccctgatatc tataacaaga
     8461 aaatatatat ataataagtt atcacgtaag tagaacatga aataacaata taattatcgt
     8521 atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc gtcattttga ctcacgcggt
     8581 cgttatagtt caaaatcagt gacacttacc gcattgacaa gcacgcctca cgggagctcc
     8641 aagcggcgac tgagatgtcc taaatgcaca gcgacggatt cgcgctattt agaaagagag
     8701 agcaatattt caagaatgca tgcgtcaatt ttacgcagac tatctttcta gggttaatct
     8761 agctgcatca ggatcatatc gtcgggtctt ttttccggct cagtcatcgc ccaagctggc
     8821 gctatctggg catcggggag gaagaagccc gtgccttttc ccgcgaggtt gaagcggcat
     8881 ggaaagagtt tgccgaggat gactgctgct gcattgacgt tgagcgaaaa cgcacgttta
     8941 ccatgatgat tcgggaaggt gtggccatgc acgcctttaa cggtgaactg ttcgttcagg
     9001 ccacctggga taccagttcg tcgcggcttt tccggacaca gttccggatg gtcagcccga
     9061 agcgcatcag caacccgaac aataccggcg acagccggaa ctgccgtgcc ggtgtgcaga
     9121 ttaatgacag cggtgcggcg ctgggatatt acgtcagcga ggacgggtat cctggctgga
     9181 tgccgcagaa atggacatgg ataccccgtg agttacccgg cgggcgcgct tggcgtaatc
     9241 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     9301 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     9361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     9421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     9481 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     9541 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     9601 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     9661 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     9721 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     9781 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     9841 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     9901 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     9961 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
    10021 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
    10081 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
    10141 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
    10201 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
    10261 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
    10321 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
    10381 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
    10441 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
    10501 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
    10561 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
    10621 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
    10681 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
    10741 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
    10801 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
    10861 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
    10921 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
    10981 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    11041 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    11101 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    11161 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    11221 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    11281 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    11341 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
    11401 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
    11461 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
    11521 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
    11581 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
    11641 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
    11701 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
    11761 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
    11821 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
    11881 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
    11941 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
    12001 acggccagtg agcgcgcctc gttcattcac gtttttgaac ccgtggagga cgggcagact
    12061 cgcggtgcaa atgtgtttta cagcgtgatg gagcagatga agatgctcga cacgctgcag
    12121 aacacgcagc tagattaacc ctagaaagat aatcatattg tgacgtacgt taaagataat
    12181 catgtgtaaa attgacgcat gtgttttatc ggtctgtata tcgaggttta tttattaatt
    12241 tgaatagata ttaagtttta ttatatttac acttacatac taataataaa ttcaacaaac
    12301 aatttattta tgtttattta tttattaaaa aaaacaaaaa ctcaaaattt cttctataaa
    12361 gtaacaaaac ttttatgagg gacagccccc ccccaaagcc cccagggatg taattacgtc
    12421 cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc cggcgctccc
    12481 cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag gtggcacggg
    12541 atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct ggggggatac
    12601 ggggaaaagg cctccacggc cgacattgat tattgactag ttattaatag taatcaatta
    12661 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
    12721 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
    12781 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
    12841 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
    12901 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
    12961 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
    13021 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
    13081 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggsgcgcs sggggggggg
    13141 gggggggggg gggggggggg gggggsgrgg sggrgaggtg cggcggcagc caatcagagc
    13201 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
    13261 cgaagcgcgc ggcgggcg
//
