LOCUS       sequence-329431-        8268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miCas9-msa fusion protein expressing plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8268)
  AUTHORS   Ma L, Ruan J, Song J, Wen L, Yang D, Zhao J, Xia X, Chen YE, Zhang 
            J, Xu J
  TITLE     MiCas9 increases large size gene knock-in rates and reduces 
            undesirable on-target and off-target indel edits.
  JOURNAL   Nat Commun. 2020 Nov 27;11(1):6082. doi: 10.1038/s41467-020-19842-2.
  PUBMED    33247137
REFERENCE   2  (bases 1 to 8268)
  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: "Nat 
            Commun."; date: "2020-11-27"; volume: "11(1):6082. doi"; pages: " 
            10.1038/s41467-020-19842-2"
COMMENT     Sequence Label: pmiCas9-msa
FEATURES             Location/Qualifiers
     source          1..8268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        473..676
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     627..647
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     718..737
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        718..736
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      744..753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(750..4151,4152..4154,4155..4751,4752,4753..4755,
                     4756..4757,4758..4760,4761..4853)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             750..4853
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             750..4853
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4860..4880
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4887..4952
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     sig_peptide     4992..5051
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             5055..5081
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5106..5447
                     /codon_start=1
                     /product="monomeric streptavidin/rhizavidin hybrid (Lim et 
                     al., 2013)"
                     /label=mSA
                     /translation="AEAGITGTWYNQSGSTFTVTAGADGNLTGQYENRAQGTGCQNSPY
                     TLTGRYNGTKLEWRVEWNNSTENCHSRTEWRGQYQGGAEARINTQWNLTYEGGSGPATE
                     QGQDTFTKVK"
     CDS             5624..5641
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5664..5681)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5670..5894
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5965..5981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5965..5981)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5978..6000)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5989..6005
                     /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(6013..6019,6020..6037,6038..6043))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6058..6079
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6196..6213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6367..6955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6447..6466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7126..7917,7918..7986))
                     /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     7749..7768
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7987..8091)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8170..8189)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
       61 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      481 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      601 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      661 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg cggccgctaa
      721 tacgactcac tatagggaga gccgccacca tggataaaaa gtattctatt ggtttagata
      781 tcggcactaa ttccgttgga tgggctgtca taaccgatga atacaaagta ccttcaaaga
      841 aatttaaggt gttggggaac acagaccgtc attcgattaa aaagaatctt atcggtgccc
      901 tcctattcga tagtggcgaa acggcagagg cgactcgcct gaaacgaacc gctcggagaa
      961 ggtatacacg tcgcaagaac cgaatatgtt acttacaaga aatttttagc aatgagatgg
     1021 ccaaagttga cgattctttc tttcaccgtt tggaagagtc cttccttgtc gaagaggaca
     1081 agaaacatga acggcacccc atctttggaa acatagtaga tgaggtggca tatcatgaaa
     1141 agtacccaac gatttatcac ctcagaaaaa agctagttga ctcaactgat aaagcggacc
     1201 tgaggttaat ctacttggct cttgcccata tgataaagtt ccgtgggcac tttctcattg
     1261 agggtgatct aaatccggac aactcggatg tcgacaaact gttcatccag ttagtacaaa
     1321 cctataatca gttgtttgaa gagaacccta taaatgcaag tggcgtggat gcgaaggcta
     1381 ttcttagcgc ccgcctctct aaatcccgac ggctagaaaa cctgatcgca caattacccg
     1441 gagagaagaa aaatgggttg ttcggtaacc ttatagcgct ctcactaggc ctgacaccaa
     1501 attttaagtc gaacttcgac ttagctgaag atgccaaatt gcagcttagt aaggacacgt
     1561 acgatgacga tctcgacaat ctactggcac aaattggaga tcagtatgcg gacttatttt
     1621 tggctgccaa aaaccttagc gatgcaatcc tcctatctga catactgaga gttaatactg
     1681 agattaccaa ggcgccgtta tccgcttcaa tgatcaaaag gtacgatgaa catcaccaag
     1741 acttgacact tctcaaggcc ctagtccgtc agcaactgcc tgagaaatat aaggaaatat
     1801 tctttgatca gtcgaaaaac gggtacgcag gttatattga cggcggagcg agtcaagagg
     1861 aattctacaa gtttatcaaa cccatattag agaagatgga tgggacggaa gagttgcttg
     1921 taaaactcaa tcgcgaagat ctactgcgaa agcagcggac tttcgacaac ggtagcattc
     1981 cacatcaaat ccacttaggc gaattgcatg ctatacttag aaggcaggag gatttttatc
     2041 cgttcctcaa agacaatcgt gaaaagattg agaaaatcct aacctttcgc ataccttact
     2101 atgtgggacc cctggcccga gggaactctc ggttcgcatg gatgacaaga aagtccgaag
     2161 aaacgattac tccatggaat tttgaggaag ttgtcgataa aggtgcgtca gctcaatcgt
     2221 tcatcgagag gatgaccaac tttgacaaga atttaccgaa cgaaaaagta ttgcctaagc
     2281 acagtttact ttacgagtat ttcacagtgt acaatgaact cacgaaagtt aagtatgtca
     2341 ctgagggcat gcgtaaaccc gcctttctaa gcggagaaca gaagaaagca atagtagatc
     2401 tgttattcaa gaccaaccgc aaagtgacag ttaagcaatt gaaagaggac tactttaaga
     2461 aaattgaatg cttcgattct gtcgagatct ccggggtaga agatcgattt aatgcgtcac
     2521 ttggtacgta tcatgacctc ctaaagataa ttaaagataa ggacttcctg gataacgaag
     2581 agaatgaaga tatcttagaa gatatagtgt tgactcttac cctctttgaa gatcgggaaa
     2641 tgattgagga aagactaaaa acatacgctc acctgttcga cgataaggtt atgaaacagt
     2701 taaagaggcg tcgctatacg ggctggggac gattgtcgcg gaaacttatc aacgggataa
     2761 gagacaagca aagtggtaaa actattctcg attttctaaa gagcgacggc ttcgccaata
     2821 ggaactttat gcagctgatc catgatgact ctttaacctt caaagaggat atacaaaagg
     2881 cacaggtttc cggacaaggg gactcattgc acgaacatat tgcgaatctt gctggttcgc
     2941 cagccatcaa aaagggcata ctccagacag tcaaagtagt ggatgagcta gttaaggtca
     3001 tgggacgtca caaaccggaa aacattgtaa tcgagatggc acgcgaaaat caaacgactc
     3061 agaaggggca aaaaaacagt cgagagcgga tgaagagaat agaagagggt attaaagaac
     3121 tgggcagcca gatcttaaag gagcatcctg tggaaaatac ccaattgcag aacgagaaac
     3181 tttacctcta ttacctacaa aatggaaggg acatgtatgt tgatcaggaa ctggacataa
     3241 accgtttatc tgattacgac gtcgatcata ttgtacccca atcctttttg aaggacgatt
     3301 caatcgacaa taaagtgctt acacgctcgg ataagaaccg agggaaaagt gacaatgttc
     3361 caagcgagga agtcgtaaag aaaatgaaga actattggcg gcagctccta aatgcgaaac
     3421 tgataacgca aagaaagttc gataacttaa ctaaagctga gaggggtggc ttgtctgaac
     3481 ttgacaaggc cggatttatt aaacgtcagc tcgtggaaac ccgccaaatc acaaagcatg
     3541 ttgcacagat actagattcc cgaatgaata cgaaatacga cgagaacgat aagctgattc
     3601 gggaagtcaa agtaatcact ttaaagtcaa aattggtgtc ggacttcaga aaggattttc
     3661 aattctataa agttagggag ataaataact accaccatgc gcacgacgct tatcttaatg
     3721 ccgtcgtagg gaccgcactc attaagaaat acccgaagct agaaagtgag tttgtgtatg
     3781 gtgattacaa agtttatgac gtccgtaaga tgatcgcgaa aagcgaacag gagataggca
     3841 aggctacagc caaatacttc ttttattcta acattatgaa tttctttaag acggaaatca
     3901 ctctggcaaa cggagagata cgcaaacgac ctttaattga aaccaatggg gagacaggtg
     3961 aaatcgtatg ggataagggc cgggacttcg cgacggtgag aaaagttttg tccatgcccc
     4021 aagtcaacat agtaaagaaa actgaggtgc agaccggagg gttttcaaag gaatcgattc
     4081 ttccaaaaag gaatagtgat aagctcatcg ctcgtaaaaa ggactgggac ccgaaaaagt
     4141 acggtggctt cgatagccct acagttgcct attctgtcct agtagtggca aaagttgaga
     4201 agggaaaatc caagaaactg aagtcagtca aagaattatt ggggataacg attatggagc
     4261 gctcgtcttt tgaaaagaac cccatcgact tccttgaggc gaaaggttac aaggaagtaa
     4321 aaaaggatct cataattaaa ctaccaaagt atagtctgtt tgagttagaa aatggccgaa
     4381 aacggatgtt ggctagcgcc ggagagcttc aaaaggggaa cgaactcgca ctaccgtcta
     4441 aatacgtgaa tttcctgtat ttagcgtccc attacgagaa gttgaaaggt tcacctgaag
     4501 ataacgaaca gaagcaactt tttgttgagc agcacaaaca ttatctcgac gaaatcatag
     4561 agcaaatttc ggaattcagt aagagagtca tcctagctga tgccaatctg gacaaagtat
     4621 taagcgcata caacaagcac agggataaac ccatacgtga gcaggcggaa aatattatcc
     4681 atttgtttac tcttaccaac ctcggcgctc cagccgcatt caagtatttt gacacaacga
     4741 tagatcgcaa acgatacact tctaccaagg aggtgctaga cgcgacactg attcaccaat
     4801 ccatcacggg attatatgaa actcggatag atttgtcaca gcttgggggt gacggatccc
     4861 ccaagaagaa gaggaaagtc tcgagcgact acaaagacca tgacggtgat tataaagatc
     4921 atgacatcga ttacaaggat gacgatgaca aggctgcagg atctggtggt tctggaggat
     4981 ctggtggttc tatggagaca gacacactcc tgctatgggt actgctgctc tgggttccag
     5041 gttccactgg tgactatcca tatgatgttc cagattatgc tggggcccag ccggccagat
     5101 ctatggcgga agcgggtatc accggcacgt ggtacaacca gtctggttct accttcaccg
     5161 ttaccgcggg tgcggacggt aacctgaccg gtcagtacga aaaccgtgcg cagggcactg
     5221 gttgccagaa ctctccgtac accctgaccg gtcgttacaa cggtaccaaa ctggaatggc
     5281 gtgttgaatg gaacaactct accgaaaact gccactctcg taccgaatgg cgtggtcagt
     5341 accagggtgg tgcggaagcg cgtatcaaca cccagtggaa cctgacctac gaaggtggtt
     5401 ctggtccggc gaccgaacag ggtcaggaca ccttcaccaa agttaaaccg tctgcggcgt
     5461 ctggatcctc tggtggttct ggaggatctg gtggttctgc tgcaggagca ttggatttcc
     5521 ttagtagact ccccttgcca cctccggtgt cccccatctg cacgtttgtg agcccagcgg
     5581 cacagaaggc atttcaaccg ccacggagtt gcgggtaacc ggtcatcatc accatcacca
     5641 ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     5701 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5761 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5821 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5881 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgatac cgtcgacctc
     5941 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6001 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctagg gtgcctaatg
     6061 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6121 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6181 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6241 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6301 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6361 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6421 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6481 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6541 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6601 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6661 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6721 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6781 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6841 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6901 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6961 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7021 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7081 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7141 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7201 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7261 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7321 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7381 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7441 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7501 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7561 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7621 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7681 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7741 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7801 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7861 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7921 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7981 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8041 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8101 ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc
     8161 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     8221 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaac
//
