LOCUS       sequence-372003-       12413 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 12413)
  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 12413)
  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_EYFP
FEATURES             Location/Qualifiers
     source          1..12413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     313..335
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     397..416
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      486..495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(492..518,519..521,522..3005,3006..3011,3012..3077,
                     3078..3080,3081..4595)
                     /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             4608..4628
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4689..4883
                     /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             4917..4937
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4947..5804
                     /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(5859..5878)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5903..5951
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6183..6394
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6404..6413
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6410..6412,6413..6415,6416..7129)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6455..6476)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6716..6735)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7063..7084
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7133..7254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7170..7189)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7224..7243
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8017..8051
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8542..8558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8542..8558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8555..8577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8566..8582
                     /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(8590..8596,8597..8614,8615..8620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8635..8656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8773..8790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8944..9532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9024..9043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9703..10494,10495..10563))
                     /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     10326..10345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10564..10668)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10694..11149)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10831..10852)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11043..11062
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11276..11298
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11290..11307
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11291..11307
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11437..11499
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11920..12299
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc
       61 attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg
      121 cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt
      181 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
      241 cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac
      301 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
      361 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
      421 ctcatcattt tggcaaagaa tctagatacc ctcgtaaagg tctagagcta gcggatcgaa
      481 cccttgccac catggacaag aagtactcca ttgggctcgc tatcggcaca aacagcgtcg
      541 gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa gttctgggca
      601 ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc gactccgggg
      661 agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga
      721 atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg gatgactctt
      781 tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac gagcgccacc
      841 caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca accatatatc
      901 atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg atctatctcg
      961 cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac ctgaacccag
     1021 acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat cagcttttcg
     1081 aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc gctaggctgt
     1141 ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag aagaacggcc
     1201 tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa tctaacttcg
     1261 acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat gatctcgaca
     1321 atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca aagaacctgt
     1381 cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc aaagctccgc
     1441 tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact ttgctgaagg
     1501 cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat cagtctaaaa
     1561 atggctacgc cggatacatt gacggcggag caagccagga ggaattttac aaatttatta
     1621 agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt aacagagaag
     1681 atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag attcacctgg
     1741 gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg aaagataaca
     1801 gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc cccctcgccc
     1861 ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc actccctgga
     1921 acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa aggatgacta
     1981 actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg ctgtacgagt
     2041 acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc
     2101 cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc aagacgaacc
     2161 ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa tgtttcgact
     2221 ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg tatcacgatc
     2281 tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag gacattcttg
     2341 aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa gaacgcttga
     2401 aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg cgccgatata
     2461 caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag cagagtggaa
     2521 agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc atgcagttga
     2581 tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt tctggccagg
     2641 gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc aaaaagggaa
     2701 tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg cataagcccg
     2761 agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga cagaagaaca
     2821 gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc caaatcctta
     2881 aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg tactacctgc
     2941 agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc tccgactacg
     3001 acgtggctgc tatcgtgccc cagtcttttc tcaaagatga ttctattgat aataaagtgt
     3061 tgacaagatc cgataaagct agagggaaga gtgataacgt cccctcagaa gaagttgtca
     3121 agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca caacggaagt
     3181 tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa gccggcttca
     3241 tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa attctcgatt
     3301 cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg aaagttatta
     3361 ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat aaggtgagag
     3421 agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta ggcactgcac
     3481 ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat aaagtgtacg
     3541 atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc gctaagtact
     3601 tcttttacag caatattatg aattttttca agaccgagat tacactggcc aatggagaga
     3661 ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg
     3721 gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa
     3781 agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa aggaacagcg
     3841 acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga ttcgattctc
     3901 ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac
     3961 tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa
     4021 accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac ctcatcatta
     4081 agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg
     4141 cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt aatttcttgt
     4201 atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag cagaagcagc
     4261 tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata agcgaattct
     4321 ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct tacaataagc
     4381 acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt actctgacca
     4441 acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga aagcggtaca
     4501 cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg gggctctatg
     4561 aaacaagaat cgacctctct cagctcggtg gagacagcag ggctgacccc aagaagaaga
     4621 ggaaggtgag tggtggagga agtggcgggt cagggtcgat ggacgcgaaa tcacttacgg
     4681 catggtcgag aacactggtt acgttcaagg acgtgtttgt ggactttaca cgtgaggagt
     4741 ggaaattgct ggatactgcg caacaaattg tgtatcgaaa tgtcatgctt gagaattaca
     4801 agaacctcgt cagtctcgga taccagttga cgaaaccgga tgtgatcctt aggctcgaaa
     4861 agggggaaga accttggctg gtatcgggag gtggttcggg tggctctgga tcaagcccaa
     4921 agaagaaacg gaaggtggaa gcctcagtgc aggtgaaaag ggtgctggaa aaatcccccg
     4981 gcaaactcct cgtgaagatg cccttccagg cttcccctgg cggaaaaggt gaagggggtg
     5041 gcgcaaccac atctgcccag gtcatggtca tcaagcgacc tggaaggaaa agaaaggccg
     5101 aggctgaccc tcaggccatt ccaaagaaac ggggacgcaa gccagggtcc gtggtcgcag
     5161 ctgcagcagc tgaggctaag aaaaaggcag tgaaggaaag ctccatccgc agtgtgcagg
     5221 agactgtcct gcccatcaag aagaggaaga ctagggagac cgtgtccatc gaggtcaaag
     5281 aagtggtcaa gcccctgctc gtgtccaccc tgggcgaaaa atctggaaag gggctcaaaa
     5341 catgcaagtc acctggacgg aaaagcaagg agtctagtcc aaaggggcgc tcaagctccg
     5401 cttctagtcc ccctaaaaag gaacaccatc accatcacca tcacgccgag tctcctaagg
     5461 ctcctatgcc actgctccca ccacctccac cacctgagcc acagtcaagc gaagacccca
     5521 tcagcccacc cgagcctcag gatctgtcct ctagtatttg caaagaggaa aagatgccca
     5581 gagcaggcag cctggagagt gatggctgtc caaaagaacc cgccaagacc cagcctatgg
     5641 tggcagccgc tgcaactacc accacaacca caactaccac agtggccgaa aaatacaagc
     5701 atcgcggcga gggcgaacga aaggacattg tgtcaagctc catgcccaga cctaaccggg
     5761 aggaaccagt cgatagtagg acacccgtga ctgagagagt ctcatagaag aattcgatcc
     5821 ctaccggtta gtaatgagtt taaacggggg aggctaactg aaacacggaa ggagacaata
     5881 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg
     5941 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     6001 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6061 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga
     6121 tctgcgaaat cggatccgcg gccgcactga ttctagctgc agaggctccg gtgcccgtca
     6181 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     6241 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     6301 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     6361 tctttttcgc aacgggtttg ccgccagaac acaggatcct cgagccacca tggtgagcaa
     6421 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     6481 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     6541 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     6601 cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     6661 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     6721 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     6781 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctgggacaca agctggagta
     6841 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     6901 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     6961 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     7021 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     7081 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag ttaacttgtt
     7141 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     7201 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     7261 ctggaattga ctcaaatgat gtcaattagt ctatcagaag ctcatctggt ctcccttccg
     7321 ggggacaaga catccctgtt taatatttaa acagcagtgt tcccaaactg ggttcttata
     7381 tcccttgctc tggtcaacca ggttgcaggg tttcctgtcc tcacaggaac gaagtcccta
     7441 aagaaacagt ggcagccagg tttagccccg gaattgactg gattcctttt ttagggccca
     7501 ttggtatggc tttttccccg tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa
     7561 gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct
     7621 gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc ggctcgctgc
     7681 tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg ggggctgtcc
     7741 ctgatatcta taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa
     7801 taacaatata attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt
     7861 cattttgact cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc
     7921 acgcctcacg ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg
     7981 cgctatttag aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta
     8041 tctttctagg gttaatctag ctgcatcagg atcatatcgt cgggtctttt ttccggctca
     8101 gtcatcgccc aagctggcgc tatctgggca tcggggagga agaagcccgt gccttttccc
     8161 gcgaggttga agcggcatgg aaagagtttg ccgaggatga ctgctgctgc attgacgttg
     8221 agcgaaaacg cacgtttacc atgatgattc gggaaggtgt ggccatgcac gcctttaacg
     8281 gtgaactgtt cgttcaggcc acctgggata ccagttcgtc gcggcttttc cggacacagt
     8341 tccggatggt cagcccgaag cgcatcagca acccgaacaa taccggcgac agccggaact
     8401 gccgtgccgg tgtgcagatt aatgacagcg gtgcggcgct gggatattac gtcagcgagg
     8461 acgggtatcc tggctggatg ccgcagaaat ggacatggat accccgtgag ttacccggcg
     8521 ggcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     8581 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     8641 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     8881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     9001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     9061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     9121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     9181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     9241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     9301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     9361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     9421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     9481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     9541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     9601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     9901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     9961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    10021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    10081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    10141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    10201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    10261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    10321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    10381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    10441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    10501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    10621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10681 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
    10741 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
    10801 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
    10861 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
    10921 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
    10981 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
    11041 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
    11101 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
    11161 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
    11221 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
    11281 tcacgacgtt gtaaaacgac ggccagtgag cgcgcctcgt tcattcacgt ttttgaaccc
    11341 gtggaggacg ggcagactcg cggtgcaaat gtgttttaca gcgtgatgga gcagatgaag
    11401 atgctcgaca cgctgcagaa cacgcagcta gattaaccct agaaagataa tcatattgtg
    11461 acgtacgtta aagataatca tgtgtaaaat tgacgcatgt gttttatcgg tctgtatatc
    11521 gaggtttatt tattaatttg aatagatatt aagttttatt atatttacac ttacatacta
    11581 ataataaatt caacaaacaa tttatttatg tttatttatt tattaaaaaa aacaaaaact
    11641 caaaatttct tctataaagt aacaaaactt ttatgaggga cagccccccc ccaaagcccc
    11701 cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc cggggctccg
    11761 ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac agcccgggca
    11821 cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct ttgagcctgc
    11881 agacacctgg ggggatacgg ggaaaaggcc tccacggccg acattgatta ttgactagtt
    11941 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
    12001 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
    12061 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
    12121 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
    12181 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
    12241 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
    12301 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
    12361 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg ggg
//
