LOCUS       sequence-371823-       13276 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9-KRAB-MeCP2 expression 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13276)
  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 13276)
  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_Puro
FEATURES             Location/Qualifiers
     source          1..13276
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          215..1231
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1155..1177
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1239..1258
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1328..1337
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1334..1360,1361..1363,1364..3847,3848..3853,3854..3919,
                     3920..3922,3923..5437)
                     /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             5450..5470
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5531..5725
                     /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             5759..5779
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5789..6646
                     /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(6701..6720)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6745..6793
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7025..7236
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             7252..7851
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7252..7271)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7748..7768
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7891..7910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7945..7964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8738..8772
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9263..9279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9263..9279)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9276..9298)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9287..9303
                     /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(9311..9317,9318..9335,9336..9341))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9356..9377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9494..9511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9665..10253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9745..9764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10424..11215,11216..11284))
                     /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     11047..11066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11285..11389)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11415..11870)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11552..11573)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11764..11783
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11997..12019
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12011..12028
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12012..12028
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   12158..12220
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        12641..13020
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tttttttwtt ttttaatttn ntwkgggggg gggggggggg gggggggggg ggggggcgcg
       61 cgcnsggssg gggsnggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
      121 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
      181 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
      241 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
      301 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
      361 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
      421 tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
      481 gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttttgc
      541 gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg
      601 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
      661 gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg
      721 cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg
      781 ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggrgggctc
      841 gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag
      901 ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg
      961 tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg
     1021 gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg
     1081 tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg
     1141 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa
     1201 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct
     1261 gtctcatcat tttggcaaag aatctagata ccctcgtaaa ggtctagagc tagcggatcg
     1321 aacccttgcc accatggaca agaagtactc cattgggctc gctatcggca caaacagcgt
     1381 cggctgggcc gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg
     1441 caataccgat cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg
     1501 ggagacggcc gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa
     1561 gaatcggatc tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc
     1621 tttcttccat aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca
     1681 cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata
     1741 tcatctgagg aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct
     1801 cgcgctggcg catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc
     1861 agacaacagc gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt
     1921 cgaagagaac ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct
     1981 gtccaaatcc cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg
     2041 cctgtttggt aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt
     2101 cgacctggcc gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga
     2161 caatctgctg gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct
     2221 gtcagacgcc attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc
     2281 gctgagcgct agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa
     2341 ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa
     2401 aaatggctac gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat
     2461 taagcccatc ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga
     2521 agatctgttg cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct
     2581 gggcgaactg cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa
     2641 cagggaaaag attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc
     2701 ccggggaaat tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg
     2761 gaacttcgag gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac
     2821 taactttgat aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga
     2881 gtacttcaca gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa
     2941 gccagcattc ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa
     3001 ccggaaagtt accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga
     3061 ctctgttgaa atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga
     3121 tctcctgaaa atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct
     3181 tgaggacatt gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt
     3241 gaaaacttac gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata
     3301 tacaggatgg gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg
     3361 aaagacaatc ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt
     3421 gatccatgat gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca
     3481 gggggacagt cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg
     3541 aatactgcag accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc
     3601 cgagaatatc gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa
     3661 cagtagggaa aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct
     3721 taaggaacac ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct
     3781 gcagaacggc agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta
     3841 cgacgtggct gctatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt
     3901 gttgacaaga tccgataaag ctagagggaa gagtgataac gtcccctcag aagaagttgt
     3961 caagaaaatg aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa
     4021 gttcgataat ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt
     4081 catcaaaagg cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga
     4141 ttcacgcatg aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat
     4201 tactctgaag tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag
     4261 agagatcaac aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc
     4321 acttatcaaa aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta
     4381 cgatgttagg aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta
     4441 cttcttttac agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga
     4501 gattcggaag cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa
     4561 gggtagggat ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa
     4621 aaagaccgaa gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag
     4681 cgacaagctg atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc
     4741 tcctacagtc gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa
     4801 actcaaaagc gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa
     4861 aaaccccatc gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat
     4921 taagcttccc aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag
     4981 tgcgggcgag ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt
     5041 gtatctggcc agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca
     5101 gctgttcgtg gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt
     5161 ctccaaaaga gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa
     5221 gcacagggat aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac
     5281 caacttgggc gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta
     5341 cacctctaca aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta
     5401 tgaaacaaga atcgacctct ctcagctcgg tggagacagc agggctgacc ccaagaagaa
     5461 gaggaaggtg agtggtggag gaagtggcgg gtcagggtcg atggacgcga aatcacttac
     5521 ggcatggtcg agaacactgg ttacgttcaa ggacgtgttt gtggacttta cacgtgagga
     5581 gtggaaattg ctggatactg cgcaacaaat tgtgtatcga aatgtcatgc ttgagaatta
     5641 caagaacctc gtcagtctcg gataccagtt gacgaaaccg gatgtgatcc ttaggctcga
     5701 aaagggggaa gaaccttggc tggtatcggg aggtggttcg ggtggctctg gatcaagccc
     5761 aaagaagaaa cggaaggtgg aagcctcagt gcaggtgaaa agggtgctgg aaaaatcccc
     5821 cggcaaactc ctcgtgaaga tgcccttcca ggcttcccct ggcggaaaag gtgaaggggg
     5881 tggcgcaacc acatctgccc aggtcatggt catcaagcga cctggaagga aaagaaaggc
     5941 cgaggctgac cctcaggcca ttccaaagaa acggggacgc aagccagggt ccgtggtcgc
     6001 agctgcagca gctgaggcta agaaaaaggc agtgaaggaa agctccatcc gcagtgtgca
     6061 ggagactgtc ctgcccatca agaagaggaa gactagggag accgtgtcca tcgaggtcaa
     6121 agaagtggtc aagcccctgc tcgtgtccac cctgggcgaa aaatctggaa aggggctcaa
     6181 aacatgcaag tcacctggac ggaaaagcaa ggagtctagt ccaaaggggc gctcaagctc
     6241 cgcttctagt ccccctaaaa aggaacacca tcaccatcac catcacgccg agtctcctaa
     6301 ggctcctatg ccactgctcc caccacctcc accacctgag ccacagtcaa gcgaagaccc
     6361 catcagccca cccgagcctc aggatctgtc ctctagtatt tgcaaagagg aaaagatgcc
     6421 cagagcaggc agcctggaga gtgatggctg tccaaaagaa cccgccaaga cccagcctat
     6481 ggtggcagcc gctgcaacta ccaccacaac cacaactacc acagtggccg aaaaatacaa
     6541 gcatcgcggc gagggcgaac gaaaggacat tgtgtcaagc tccatgccca gacctaaccg
     6601 ggaggaacca gtcgatagta ggacacccgt gactgagaga gtctcataga agaattcgat
     6661 ccctaccggt tagtaatgag tttaaacggg ggaggctaac tgaaacacgg aaggagacaa
     6721 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     6781 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     6841 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     6901 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     6961 gatctgcgaa atcggatccg cggccgcact gattctagct gcagaggctc cggtgcccgt
     7021 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     7081 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     7141 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     7201 gttctttttc gcaacgggtt tgccgccaga acacaggatc ctcgagccac catgaccgag
     7261 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     7321 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     7381 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     7441 gtgtgggtcg cggacgacgg cgccgccgtg gcggtctgga ccacgccgga gagcgtcgaa
     7501 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     7561 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     7621 ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     7681 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     7741 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     7801 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta agttaacttt
     7861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     7921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     7981 tctggaattg actcaaatga tgtcaattag tctatcagaa gctcatctgg tctcccttcc
     8041 gggggacaag acatccctgt ttaatattta aacagcagtg ttcccaaact gggttcttat
     8101 atcccttgct ctggtcaacc aggttgcagg gtttcctgtc ctcacaggaa cgaagtccct
     8161 aaagaaacag tggcagccag gtttagcccc ggaattgact ggattccttt tttagggccc
     8221 attggtatgg ctttttcccc gtatcccccc aggtgtctgc aggctcaaag agcagcgaga
     8281 agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt ccccgcacgc
     8341 tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg cggctcgctg
     8401 ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg gggggctgtc
     8461 cctgatatct ataacaagaa aatatatata taataagtta tcacgtaagt agaacatgaa
     8521 ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg
     8581 tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg cattgacaag
     8641 cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag cgacggattc
     8701 gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt tacgcagact
     8761 atctttctag ggttaatcta gctgcatcag gatcatatcg tcgggtcttt tttccggctc
     8821 agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg tgccttttcc
     8881 cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg cattgacgtt
     8941 gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca cgcctttaac
     9001 ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt ccggacacag
     9061 ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga cagccggaac
     9121 tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta cgtcagcgag
     9181 gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga gttacccggc
     9241 gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     9301 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     9361 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     9421 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     9481 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     9541 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     9601 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     9661 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     9721 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     9781 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     9841 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     9901 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     9961 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    10021 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    10081 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    10141 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    10201 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
    10261 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    10321 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
    10381 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
    10441 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
    10501 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
    10561 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    10621 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    10681 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    10741 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    10801 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    10861 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    10921 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    10981 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    11041 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    11101 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    11161 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    11221 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    11281 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    11341 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    11401 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
    11461 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
    11521 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
    11581 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
    11641 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
    11701 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
    11761 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
    11821 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
    11881 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
    11941 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
    12001 gtcacgacgt tgtaaaacga cggccagtga gcgcgcctcg ttcattcacg tttttgaacc
    12061 cgtggaggac gggcagactc gcggtgcaaa tgtgttttac agcgtgatgg agcagatgaa
    12121 gatgctcgac acgctgcaga acacgcagct agattaaccc tagaaagata atcatattgt
    12181 gacgtacgtt aaagataatc atgtgtaaaa ttgacgcatg tgttttatcg gtctgtatat
    12241 cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca cttacatact
    12301 aataataaat tcaacaaaca atttatttat gtttatttat ttattaaaaa aaacaaaaac
    12361 tcaaaatttc ttctataaag taacaaaact tttatgaggg acagcccccc cccaaagccc
    12421 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
    12481 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
    12541 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
    12601 cagacacctg gggggatacg gggaaaaggc ctccacggcc gacattgatt attgactagt
    12661 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
    12721 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
    12781 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
    12841 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
    12901 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
    12961 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
    13021 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
    13081 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
    13141 ggssssscsc nggcgggggg gggggggggg gggggggggg gggggggggg gggggggggg
    13201 gggggggggs gssskngnnn gggsgggggg ggggggggrg gggggggkgg gggggggggg
    13261 gggggggggg gggggg
//
