LOCUS       sequence-371892-       12271 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 12271)
  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 12271)
  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_EBFP2
FEATURES             Location/Qualifiers
     source          1..12271
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     258..280
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     342..361
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      431..440
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(437..463,464..466,467..2950,2951..2956,2957..3022,
                     3023..3025,3026..4540)
                     /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             4553..4573
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4634..4828
                     /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             4862..4882
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4892..5749
                     /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(5804..5823)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5848..5896
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6128..6339
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6349..6358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6355..6357,6358..6360,6361..7074)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(6400..6421)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7114..7133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7168..7187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7961..7995
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8486..8502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8486..8502)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8499..8521)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8510..8526
                     /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(8534..8540,8541..8558,8559..8564))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8579..8600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8717..8734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8888..9476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8968..8987)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9647..10438,10439..10507))
                     /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     10270..10289
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10508..10612)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10638..11093)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10775..10796)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10987..11006
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11220..11242
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11234..11251
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11235..11251
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11381..11443
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11864..12243
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat
       61 ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa
      121 gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg
      181 ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg
      241 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc
      301 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
      361 gctgtctcat cattttggca aagaatctag ataccctcgt aaaggtctag agctagcgga
      421 tcgaaccctt gccaccatgg acaagaagta ctccattggg ctcgctatcg gcacaaacag
      481 cgtcggctgg gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct
      541 gggcaatacc gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc
      601 cggggagacg gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag
      661 aaagaatcgg atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga
      721 ctctttcttc cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg
      781 ccacccaatc tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat
      841 atatcatctg aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta
      901 tctcgcgctg gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa
      961 cccagacaac agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct
     1021 tttcgaagag aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag
     1081 gctgtccaaa tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa
     1141 cggcctgttt ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa
     1201 cttcgacctg gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct
     1261 cgacaatctg ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa
     1321 cctgtcagac gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc
     1381 tccgctgagc gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct
     1441 gaaggccctt gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc
     1501 taaaaatggc tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt
     1561 tattaagccc atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag
     1621 agaagatctg ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca
     1681 cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga
     1741 taacagggaa aagattgaga aaatcctcac atttcggata ccctactatg taggccccct
     1801 cgcccgggga aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc
     1861 ctggaacttc gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat
     1921 gactaacttt gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta
     1981 cgagtacttc acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag
     2041 aaagccagca ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac
     2101 gaaccggaaa gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt
     2161 cgactctgtt gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca
     2221 cgatctcctg aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat
     2281 tcttgaggac attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg
     2341 cttgaaaact tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg
     2401 atatacagga tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag
     2461 tggaaagaca atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca
     2521 gttgatccat gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg
     2581 ccagggggac agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa
     2641 gggaatactg cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa
     2701 gcccgagaat atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa
     2761 gaacagtagg gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat
     2821 ccttaaggaa cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta
     2881 cctgcagaac ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga
     2941 ctacgacgtg gctgctatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa
     3001 agtgttgaca agatccgata aagctagagg gaagagtgat aacgtcccct cagaagaagt
     3061 tgtcaagaaa atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg
     3121 gaagttcgat aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg
     3181 cttcatcaaa aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct
     3241 cgattcacgc atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt
     3301 tattactctg aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt
     3361 gagagagatc aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac
     3421 tgcacttatc aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt
     3481 gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa
     3541 gtacttcttt tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg
     3601 agagattcgg aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga
     3661 caagggtagg gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt
     3721 taaaaagacc gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa
     3781 cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga
     3841 ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa
     3901 aaaactcaaa agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga
     3961 aaaaaacccc atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat
     4021 cattaagctt cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc
     4081 tagtgcgggc gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt
     4141 cttgtatctg gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa
     4201 gcagctgttc gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga
     4261 attctccaaa agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa
     4321 taagcacagg gataagccca tcagggagca ggcagaaaac attatccact tgtttactct
     4381 gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg
     4441 gtacacctct acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct
     4501 ctatgaaaca agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa
     4561 gaagaggaag gtgagtggtg gaggaagtgg cgggtcaggg tcgatggacg cgaaatcact
     4621 tacggcatgg tcgagaacac tggttacgtt caaggacgtg tttgtggact ttacacgtga
     4681 ggagtggaaa ttgctggata ctgcgcaaca aattgtgtat cgaaatgtca tgcttgagaa
     4741 ttacaagaac ctcgtcagtc tcggatacca gttgacgaaa ccggatgtga tccttaggct
     4801 cgaaaagggg gaagaacctt ggctggtatc gggaggtggt tcgggtggct ctggatcaag
     4861 cccaaagaag aaacggaagg tggaagcctc agtgcaggtg aaaagggtgc tggaaaaatc
     4921 ccccggcaaa ctcctcgtga agatgccctt ccaggcttcc cctggcggaa aaggtgaagg
     4981 gggtggcgca accacatctg cccaggtcat ggtcatcaag cgacctggaa ggaaaagaaa
     5041 ggccgaggct gaccctcagg ccattccaaa gaaacgggga cgcaagccag ggtccgtggt
     5101 cgcagctgca gcagctgagg ctaagaaaaa ggcagtgaag gaaagctcca tccgcagtgt
     5161 gcaggagact gtcctgccca tcaagaagag gaagactagg gagaccgtgt ccatcgaggt
     5221 caaagaagtg gtcaagcccc tgctcgtgtc caccctgggc gaaaaatctg gaaaggggct
     5281 caaaacatgc aagtcacctg gacggaaaag caaggagtct agtccaaagg ggcgctcaag
     5341 ctccgcttct agtcccccta aaaaggaaca ccatcaccat caccatcacg ccgagtctcc
     5401 taaggctcct atgccactgc tcccaccacc tccaccacct gagccacagt caagcgaaga
     5461 ccccatcagc ccacccgagc ctcaggatct gtcctctagt atttgcaaag aggaaaagat
     5521 gcccagagca ggcagcctgg agagtgatgg ctgtccaaaa gaacccgcca agacccagcc
     5581 tatggtggca gccgctgcaa ctaccaccac aaccacaact accacagtgg ccgaaaaata
     5641 caagcatcgc ggcgagggcg aacgaaagga cattgtgtca agctccatgc ccagacctaa
     5701 ccgggaggaa ccagtcgata gtaggacacc cgtgactgag agagtctcat agaagaattc
     5761 gatccctacc ggttagtaat gagtttaaac gggggaggct aactgaaaca cggaaggaga
     5821 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     5881 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5941 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     6001 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     6061 gcagatctgc gaaatcggat ccgcggccgc actgattcta gctgcagagg ctccggtgcc
     6121 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     6181 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     6241 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     6301 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg atcctcgagc caccatggtg
     6361 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     6421 gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac caacggcaag
     6481 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     6541 accaccctga gccacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     6601 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     6661 gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctagtgaac
     6721 cgcatcgagc tgaagggcgt cgacttcaag gaggacggca acatcctggg gcacaagctg
     6781 gagtacaact tcaacagcca caacatctat atcatggccg tcaagcagaa gaacggcatc
     6841 aaggtgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac
     6901 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacag ccactacctg
     6961 agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     7021 gagttccgca ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagttaac
     7081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7201 atgtctggaa ttgactcaaa tgatgtcaat tagtctatca gaagctcatc tggtctccct
     7261 tccgggggac aagacatccc tgtttaatat ttaaacagca gtgttcccaa actgggttct
     7321 tatatccctt gctctggtca accaggttgc agggtttcct gtcctcacag gaacgaagtc
     7381 cctaaagaaa cagtggcagc caggtttagc cccggaattg actggattcc ttttttaggg
     7441 cccattggta tggctttttc cccgtatccc cccaggtgtc tgcaggctca aagagcagcg
     7501 agaagcgttc agaggaaagc gatcccgtgc caccttcccc gtgcccgggc tgtccccgca
     7561 cgctgccggc tcggggatgc ggggggagcg ccggaccgga gcggagcccc gggcggctcg
     7621 ctgctgcccc ctagcggggg agggacgtaa ttacatccct gggggctttg ggggggggct
     7681 gtccctgata tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat
     7741 gaaataacaa tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat
     7801 gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac
     7861 aagcacgcct cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga
     7921 ttcgcgctat ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag
     7981 actatctttc tagggttaat ctagctgcat caggatcata tcgtcgggtc ttttttccgg
     8041 ctcagtcatc gcccaagctg gcgctatctg ggcatcgggg aggaagaagc ccgtgccttt
     8101 tcccgcgagg ttgaagcggc atggaaagag tttgccgagg atgactgctg ctgcattgac
     8161 gttgagcgaa aacgcacgtt taccatgatg attcgggaag gtgtggccat gcacgccttt
     8221 aacggtgaac tgttcgttca ggccacctgg gataccagtt cgtcgcggct tttccggaca
     8281 cagttccgga tggtcagccc gaagcgcatc agcaacccga acaataccgg cgacagccgg
     8341 aactgccgtg ccggtgtgca gattaatgac agcggtgcgg cgctgggata ttacgtcagc
     8401 gaggacgggt atcctggctg gatgccgcag aaatggacat ggataccccg tgagttaccc
     8461 ggcgggcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     8521 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     8581 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     8641 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     8701 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     8761 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     8821 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     8881 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     8941 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     9001 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     9061 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     9121 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     9181 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     9241 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     9301 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     9361 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     9421 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     9481 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     9541 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     9601 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     9661 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     9721 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     9781 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     9841 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     9901 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9961 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
    10021 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
    10081 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
    10141 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    10201 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    10261 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    10321 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    10381 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    10441 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    10501 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    10561 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    10621 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
    10681 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
    10741 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
    10801 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
    10861 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
    10921 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
    10981 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
    11041 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
    11101 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
    11161 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
    11221 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcc tcgttcattc acgtttttga
    11281 acccgtggag gacgggcaga ctcgcggtgc aaatgtgttt tacagcgtga tggagcagat
    11341 gaagatgctc gacacgctgc agaacacgca gctagattaa ccctagaaag ataatcatat
    11401 tgtgacgtac gttaaagata atcatgtgta aaattgacgc atgtgtttta tcggtctgta
    11461 tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt acacttacat
    11521 actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa aaaaaacaaa
    11581 aactcaaaat ttcttctata aagtaacaaa acttttatga gggacagccc ccccccaaag
    11641 cccccaggga tgtaattacg tccctccccc gctagggggc agcagcgagc cgcccggggc
    11701 tccgctccgg tccggcgctc cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg
    11761 ggcacgggga aggtggcacg ggatcgcttt cctctgaacg cttctcgctg ctctttgagc
    11821 ctgcagacac ctggggggat acggggaaaa ggcctccacg gccgacattg attattgact
    11881 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
    11941 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
    12001 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
    12061 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
    12121 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
    12181 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcmt cgctattacc
    12241 atggtcgagg tgagccccac gttctgcttc a
//
