LOCUS       sequence-354070-       11851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11851)
  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 11851)
  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_Puro
FEATURES             Location/Qualifiers
     source          1..11851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          122..1138
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1062..1084
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1146..1165
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1235..1244
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1241..1267,1268..1270,1271..3754,3755..3760,3761..3826,
                     3827..3829,3830..5344)
                     /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             5357..5377
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5402..5421)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5446..5494
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5726..5937
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5953..6552
                     /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(5953..5972)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6449..6469
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6571..6692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6608..6627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6662..6681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7455..7489
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7980..7996)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7980..7996)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7993..8015)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8004..8020
                     /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(8028..8034,8035..8052,8053..8058))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8073..8094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8211..8228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8382..8970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8462..8481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9141..9932,9933..10001))
                     /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     9764..9783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10002..10106)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10132..10587)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10269..10290)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10481..10500
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10714..10736
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10728..10745
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10729..10745
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   10875..10937
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11358..11737
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
       61 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      121 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      181 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      241 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      301 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      361 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      421 ctgcgggcgc ggcgcggggc tttttgcgct ccgcagtgtg cgcgagggga gcgcggccgg
      481 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
      541 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
      601 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
      661 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
      721 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
      781 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
      841 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
      901 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
      961 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1021 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1081 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1141 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat ctagataccc
     1201 tcgtaaaggt ctagagctag cggatcgaac ccttgccacc atggacaaga agtactccat
     1261 tgggctcgct atcggcacaa acagcgtcgg ctgggccgtc attacggacg agtacaaggt
     1321 gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct
     1381 cattggcgcc ctcctgttcg actccgggga gacggccgaa gccacgcggc tcaaaagaac
     1441 agcacggcgc agatataccc gcagaaagaa tcggatctgc tacctgcagg agatctttag
     1501 taatgagatg gctaaggtgg atgactcttt cttccatagg ctggaggagt cctttttggt
     1561 ggaggaggat aaaaagcacg agcgccaccc aatctttggc aatatcgtgg acgaggtggc
     1621 gtaccatgaa aagtacccaa ccatatatca tctgaggaag aagcttgtag acagtactga
     1681 taaggctgac ttgcggttga tctatctcgc gctggcgcat atgatcaaat ttcggggaca
     1741 cttcctcatc gagggggacc tgaacccaga caacagcgat gtcgacaaac tctttatcca
     1801 actggttcag acttacaatc agcttttcga agagaacccg atcaacgcat ccggagttga
     1861 cgccaaagca atcctgagcg ctaggctgtc caaatcccgg cggctcgaaa acctcatcgc
     1921 acagctccct ggggagaaga agaacggcct gtttggtaat cttatcgccc tgtcactcgg
     1981 gctgaccccc aactttaaat ctaacttcga cctggccgaa gatgccaagc ttcaactgag
     2041 caaagacacc tacgatgatg atctcgacaa tctgctggcc cagatcggcg accagtacgc
     2101 agaccttttt ttggcggcaa agaacctgtc agacgccatt ctgctgagtg atattctgcg
     2161 agtgaacacg gagatcacca aagctccgct gagcgctagt atgatcaagc gctatgatga
     2221 gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga cagcaactgc ctgagaagta
     2281 caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc ggatacattg acggcggagc
     2341 aagccaggag gaattttaca aatttattaa gcccatcttg gaaaaaatgg acggcaccga
     2401 ggagctgctg gtaaagctta acagagaaga tctgttgcgc aaacagcgca ctttcgacaa
     2461 tggaagcatc ccccaccaga ttcacctggg cgaactgcac gctatcctca ggcggcaaga
     2521 ggatttctac ccctttttga aagataacag ggaaaagatt gagaaaatcc tcacatttcg
     2581 gataccctac tatgtaggcc ccctcgcccg gggaaattcc agattcgcgt ggatgactcg
     2641 caaatcagaa gagaccatca ctccctggaa cttcgaggaa gtcgtggata agggggcctc
     2701 tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa aatctgccta acgaaaaggt
     2761 gcttcctaaa cactctctgc tgtacgagta cttcacagtt tataacgagc tcaccaaggt
     2821 caaatacgtc acagaaggga tgagaaagcc agcattcctg tctggagagc agaagaaagc
     2881 tatcgtggac ctcctcttca agacgaaccg gaaagttacc gtgaaacagc tcaaagaaga
     2941 ctatttcaaa aagattgaat gtttcgactc tgttgaaatc agcggagtgg aggatcgctt
     3001 caacgcatcc ctgggaacgt atcacgatct cctgaaaatc attaaagaca aggacttcct
     3061 ggacaatgag gagaacgagg acattcttga ggacattgtc ctcaccctta cgttgtttga
     3121 agatagggag atgattgaag aacgcttgaa aacttacgct catctcttcg acgacaaagt
     3181 catgaaacag ctcaagaggc gccgatatac aggatggggg cggctgtcaa gaaaactgat
     3241 caatgggatc cgagacaagc agagtggaaa gacaatcctg gattttctta agtccgatgg
     3301 atttgccaac cggaacttca tgcagttgat ccatgatgac tctctcacct ttaaggagga
     3361 catccagaaa gcacaagttt ctggccaggg ggacagtctt cacgagcaca tcgctaatct
     3421 tgcaggtagc ccagctatca aaaagggaat actgcagacc gttaaggtcg tggatgaact
     3481 cgtcaaagta atgggaaggc ataagcccga gaatatcgtt atcgagatgg cccgagagaa
     3541 ccaaactacc cagaagggac agaagaacag tagggaaagg atgaagagga ttgaagaggg
     3601 tataaaagaa ctggggtccc aaatccttaa ggaacaccca gttgaaaaca cccagcttca
     3661 gaatgagaag ctctacctgt actacctgca gaacggcagg gacatgtacg tggatcagga
     3721 actggacatc aatcggctct ccgactacga cgtggctgct atcgtgcccc agtcttttct
     3781 caaagatgat tctattgata ataaagtgtt gacaagatcc gataaagcta gagggaagag
     3841 tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa aattattggc ggcagctgct
     3901 gaacgccaaa ctgatcacac aacggaagtt cgataatctg actaaggctg aacgaggtgg
     3961 cctgtctgag ttggataaag ccggcttcat caaaaggcag cttgttgaga cacgccagat
     4021 caccaagcac gtggcccaaa ttctcgattc acgcatgaac accaagtacg atgaaaatga
     4081 caaactgatt cgagaggtga aagttattac tctgaagtct aagctggtct cagatttcag
     4141 aaaggacttt cagttttata aggtgagaga gatcaacaat taccaccatg cgcatgatgc
     4201 ctacctgaat gcagtggtag gcactgcact tatcaaaaaa tatcccaagc ttgaatctga
     4261 atttgtttac ggagactata aagtgtacga tgttaggaaa atgatcgcaa agtctgagca
     4321 ggaaataggc aaggccaccg ctaagtactt cttttacagc aatattatga attttttcaa
     4381 gaccgagatt acactggcca atggagagat tcggaagcga ccacttatcg aaacaaacgg
     4441 agaaacagga gaaatcgtgt gggacaaggg tagggatttc gcgacagtcc ggaaggtcct
     4501 gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta cagaccggag gcttctccaa
     4561 ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc gcacgcaaaa aagattggga
     4621 ccccaagaaa tacggcggat tcgattctcc tacagtcgct tacagtgtac tggttgtggc
     4681 caaagtggag aaagggaagt ctaaaaaact caaaagcgtc aaggaactgc tgggcatcac
     4741 aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac tttctcgagg cgaaaggata
     4801 taaagaggtc aaaaaagacc tcatcattaa gcttcccaag tactctctct ttgagcttga
     4861 aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg cagaaaggta acgagctggc
     4921 actgccctct aaatacgtta atttcttgta tctggccagc cactatgaaa agctcaaagg
     4981 gtctcccgaa gataatgagc agaagcagct gttcgtggaa caacacaaac actaccttga
     5041 tgagatcatc gagcaaataa gcgaattctc caaaagagtg atcctcgccg acgctaacct
     5101 cgataaggtg ctttctgctt acaataagca cagggataag cccatcaggg agcaggcaga
     5161 aaacattatc cacttgttta ctctgaccaa cttgggcgcg cctgcagcct tcaagtactt
     5221 cgacaccacc atagacagaa agcggtacac ctctacaaag gaggtcctgg acgccacact
     5281 gattcatcag tcaattacgg ggctctatga aacaagaatc gacctctctc agctcggtgg
     5341 agacagcagg gctgacccca agaagaagag gaaggtgtga gtttaaacgg gggaggctaa
     5401 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5461 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     5521 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     5581 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5641 gcggcaggcc ctgccatagc agatctgcga aatcggatcc gcggccgcac tgattctagc
     5701 tgcagaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     5761 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     5821 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     5881 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggat
     5941 cctcgagcca ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     6001 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     6061 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     6121 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     6181 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     6241 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     6301 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     6361 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     6421 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     6481 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     6541 cccggtgcct gaatctagtg tcgacaagtt aacttgttta ttgcagctta taatggttac
     6601 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6661 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggaattgact caaatgatgt
     6721 caattagtct atcagaagct catctggtct cccttccggg ggacaagaca tccctgttta
     6781 atatttaaac agcagtgttc ccaaactggg ttcttatatc ccttgctctg gtcaaccagg
     6841 ttgcagggtt tcctgtcctc acaggaacga agtccctaaa gaaacagtgg cagccaggtt
     6901 tagccccgga attgactgga ttcctttttt agggcccatt ggtatggctt tttccccgta
     6961 tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc
     7021 gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg
     7081 agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac
     7141 gtaattacat ccctgggggc tttggggggg ggctgtccct gatatctata acaagaaaat
     7201 atatatataa taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga
     7261 gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt
     7321 atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc
     7381 ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca
     7441 atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt taatctagct
     7501 gcatcaggat catatcgtcg ggtctttttt ccggctcagt catcgcccaa gctggcgcta
     7561 tctgggcatc ggggaggaag aagcccgtgc cttttcccgc gaggttgaag cggcatggaa
     7621 agagtttgcc gaggatgact gctgctgcat tgacgttgag cgaaaacgca cgtttaccat
     7681 gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt gaactgttcg ttcaggccac
     7741 ctgggatacc agttcgtcgc ggcttttccg gacacagttc cggatggtca gcccgaagcg
     7801 catcagcaac ccgaacaata ccggcgacag ccggaactgc cgtgccggtg tgcagattaa
     7861 tgacagcggt gcggcgctgg gatattacgt cagcgaggac gggtatcctg gctggatgcc
     7921 gcagaaatgg acatggatac cccgtgagtt acccggcggg cgcgcttggc gtaatcatgg
     7981 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     8041 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     8101 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     8161 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     8221 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     8281 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     8341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8821 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     9001 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     9061 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     9121 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     9181 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9241 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     9301 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     9361 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9421 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9481 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9541 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9601 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9661 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9721 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9781 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9841 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9901 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9961 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    10021 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    10081 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
    10141 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
    10201 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
    10261 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
    10321 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
    10381 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
    10441 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
    10501 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
    10561 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
    10621 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
    10681 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
    10741 ccagtgagcg cgcctcgttc attcacgttt ttgaacccgt ggaggacggg cagactcgcg
    10801 gtgcaaatgt gttttacagc gtgatggagc agatgaagat gctcgacacg ctgcagaaca
    10861 cgcagctaga ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg
    10921 tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa
    10981 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt
    11041 tatttatgtt tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa
    11101 caaaactttt atgagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc
    11161 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg
    11221 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg
    11281 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg
    11341 aaaaggcctc cacggccgac attgattatt gactagttat taatagtaat caattacggg
    11401 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
    11461 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
    11521 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
    11581 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
    11641 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
    11701 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
    11761 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
    11821 attattttgt gcagcgatgg gggcgggggg g
//
