LOCUS       sequence-354111-       12109 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 12109)
  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 12109)
  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_Blasto
FEATURES             Location/Qualifiers
     source          1..12109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     321..343
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     405..424
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      494..503
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(500..526,527..529,530..3013,3014..3019,3020..3085,
                     3086..3088,3089..4603)
                     /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             4616..4636
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4697..4891
                     /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             4925..4945
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4955..5812
                     /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(5867..5886)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5911..5959
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6191..6402
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6412..6421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6418..6816
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    6820..6941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6857..6876)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6911..6930
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7704..7738
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8229..8245)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8229..8245)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8242..8264)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8253..8269
                     /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(8277..8283,8284..8301,8302..8307))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8322..8343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8460..8477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8631..9219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8711..8730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9390..10181,10182..10250))
                     /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     10013..10032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10251..10355)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10381..10836)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10518..10539)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10730..10749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10963..10985
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10977..10994
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10978..10994
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11124..11186
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11607..11986
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
       61 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
      121 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
      181 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
      241 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
      301 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
      361 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
      421 tgtgctgtct catcattttg gcaaagaatc tagataccct cgtaaaggtc tagagctagc
      481 ggatcgaacc cttgccacca tggacaagaa gtactccatt gggctcgcta tcggcacaaa
      541 cagcgtcggc tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt
      601 tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc tcctgttcga
      661 ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca gatatacccg
      721 cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg ctaaggtgga
      781 tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata aaaagcacga
      841 gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa agtacccaac
      901 catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact tgcggttgat
      961 ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg agggggacct
     1021 gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga cttacaatca
     1081 gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa tcctgagcgc
     1141 taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg gggagaagaa
     1201 gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca actttaaatc
     1261 taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct acgatgatga
     1321 tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt tggcggcaaa
     1381 gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg agatcaccaa
     1441 agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag acttgacttt
     1501 gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt tcttcgatca
     1561 gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg aattttacaa
     1621 atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg taaagcttaa
     1681 cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc cccaccagat
     1741 tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc cctttttgaa
     1801 agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact atgtaggccc
     1861 cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag agaccatcac
     1921 tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct tcatcgaaag
     1981 gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac actctctgct
     2041 gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca cagaagggat
     2101 gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc tcctcttcaa
     2161 gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa agattgaatg
     2221 tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc tgggaacgta
     2281 tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg agaacgagga
     2341 cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga tgattgaaga
     2401 acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc tcaagaggcg
     2461 ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc gagacaagca
     2521 gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc ggaacttcat
     2581 gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag cacaagtttc
     2641 tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc cagctatcaa
     2701 aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca
     2761 taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc agaagggaca
     2821 gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac tggggtccca
     2881 aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc tctacctgta
     2941 ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca atcggctctc
     3001 cgactacgac gtggctgcta tcgtgcccca gtcttttctc aaagatgatt ctattgataa
     3061 taaagtgttg acaagatccg ataaagctag agggaagagt gataacgtcc cctcagaaga
     3121 agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac tgatcacaca
     3181 acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt tggataaagc
     3241 cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg tggcccaaat
     3301 tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc gagaggtgaa
     3361 agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc agttttataa
     3421 ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg cagtggtagg
     3481 cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg gagactataa
     3541 agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca aggccaccgc
     3601 taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta cactggccaa
     3661 tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag aaatcgtgtg
     3721 ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc aggtgaacat
     3781 cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc tcccgaaaag
     3841 gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat acggcggatt
     3901 cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga aagggaagtc
     3961 taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc gatcaagctt
     4021 cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca aaaaagacct
     4081 catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga aacgaatgct
     4141 cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta aatacgttaa
     4201 tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag ataatgagca
     4261 gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg agcaaataag
     4321 cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc tttctgctta
     4381 caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc acttgtttac
     4441 tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca tagacagaaa
     4501 gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt caattacggg
     4561 gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg ctgaccccaa
     4621 gaagaagagg aaggtgagtg gtggaggaag tggcgggtca gggtcgatgg acgcgaaatc
     4681 acttacggca tggtcgagaa cactggttac gttcaaggac gtgtttgtgg actttacacg
     4741 tgaggagtgg aaattgctgg atactgcgca acaaattgtg tatcgaaatg tcatgcttga
     4801 gaattacaag aacctcgtca gtctcggata ccagttgacg aaaccggatg tgatccttag
     4861 gctcgaaaag ggggaagaac cttggctggt atcgggaggt ggttcgggtg gctctggatc
     4921 aagcccaaag aagaaacgga aggtggaagc ctcagtgcag gtgaaaaggg tgctggaaaa
     4981 atcccccggc aaactcctcg tgaagatgcc cttccaggct tcccctggcg gaaaaggtga
     5041 agggggtggc gcaaccacat ctgcccaggt catggtcatc aagcgacctg gaaggaaaag
     5101 aaaggccgag gctgaccctc aggccattcc aaagaaacgg ggacgcaagc cagggtccgt
     5161 ggtcgcagct gcagcagctg aggctaagaa aaaggcagtg aaggaaagct ccatccgcag
     5221 tgtgcaggag actgtcctgc ccatcaagaa gaggaagact agggagaccg tgtccatcga
     5281 ggtcaaagaa gtggtcaagc ccctgctcgt gtccaccctg ggcgaaaaat ctggaaaggg
     5341 gctcaaaaca tgcaagtcac ctggacggaa aagcaaggag tctagtccaa aggggcgctc
     5401 aagctccgct tctagtcccc ctaaaaagga acaccatcac catcaccatc acgccgagtc
     5461 tcctaaggct cctatgccac tgctcccacc acctccacca cctgagccac agtcaagcga
     5521 agaccccatc agcccacccg agcctcagga tctgtcctct agtatttgca aagaggaaaa
     5581 gatgcccaga gcaggcagcc tggagagtga tggctgtcca aaagaacccg ccaagaccca
     5641 gcctatggtg gcagccgctg caactaccac cacaaccaca actaccacag tggccgaaaa
     5701 atacaagcat cgcggcgagg gcgaacgaaa ggacattgtg tcaagctcca tgcccagacc
     5761 taaccgggag gaaccagtcg atagtaggac acccgtgact gagagagtct catagaagaa
     5821 ttcgatccct accggttagt aatgagttta aacgggggag gctaactgaa acacggaagg
     5881 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5941 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     6001 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     6061 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     6121 atagcagatc tgcgaaatcg gatccgcggc cgcactgatt ctagctgcag aggctccggt
     6181 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     6241 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     6301 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     6361 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggatcctcg agccaccatg
     6421 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     6481 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     6541 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     6601 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     6661 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     6721 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     6781 gaattgctgc cctctggtta tgtgtgggag ggctaagtta acttgtttat tgcagcttat
     6841 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6901 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gaattgactc
     6961 aaatgatgtc aattagtcta tcagaagctc atctggtctc ccttccgggg gacaagacat
     7021 ccctgtttaa tatttaaaca gcagtgttcc caaactgggt tcttatatcc cttgctctgg
     7081 tcaaccaggt tgcagggttt cctgtcctca caggaacgaa gtccctaaag aaacagtggc
     7141 agccaggttt agccccggaa ttgactggat tcctttttta gggcccattg gtatggcttt
     7201 ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa
     7261 agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga
     7321 tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc cccctagcgg
     7381 gggagggacg taattacatc cctgggggct ttgggggggg gctgtccctg atatctataa
     7441 caagaaaata tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt
     7501 atcgtatgag ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac
     7561 gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga
     7621 gctccaagcg gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa
     7681 gagagagcaa tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt
     7741 aatctagctg catcaggatc atatcgtcgg gtcttttttc cggctcagtc atcgcccaag
     7801 ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc ttttcccgcg aggttgaagc
     7861 ggcatggaaa gagtttgccg aggatgactg ctgctgcatt gacgttgagc gaaaacgcac
     7921 gtttaccatg atgattcggg aaggtgtggc catgcacgcc tttaacggtg aactgttcgt
     7981 tcaggccacc tgggatacca gttcgtcgcg gcttttccgg acacagttcc ggatggtcag
     8041 cccgaagcgc atcagcaacc cgaacaatac cggcgacagc cggaactgcc gtgccggtgt
     8101 gcagattaat gacagcggtg cggcgctggg atattacgtc agcgaggacg ggtatcctgg
     8161 ctggatgccg cagaaatgga catggatacc ccgtgagtta cccggcgggc gcgcttggcg
     8221 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     8281 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     8341 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     8401 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     8461 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     8521 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     8581 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     8641 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     8701 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     8761 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     8821 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8881 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8941 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     9001 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     9061 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     9121 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     9181 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     9241 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     9301 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     9361 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     9421 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     9481 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     9541 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     9601 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     9661 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     9721 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     9781 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9841 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9901 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9961 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    10021 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    10081 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    10141 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    10201 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    10261 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    10321 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    10381 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
    10441 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
    10501 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
    10561 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
    10621 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
    10681 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
    10741 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
    10801 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
    10861 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
    10921 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
    10981 aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg gaggacgggc
    11041 agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg ctcgacacgc
    11101 tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg tacgttaaag
    11161 ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag gtttatttat
    11221 taatttgaat agatattaag ttttattata tttacactta catactaata ataaattcaa
    11281 caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa aatttcttct
    11341 ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag ggatgtaatt
    11401 acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc cggtccggcg
    11461 ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc
    11521 acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga cacctggggg
    11581 gatacgggga aaaggcctcc acggccgaca ttgattattg actagttatt aatagtaatc
    11641 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
    11701 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
    11761 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
    11821 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
    11881 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
    11941 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
    12001 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
    12061 tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggggg
//
