LOCUS       sequence-241051-        9444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dead Cas9 (dCas9)-CBE4-gam.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9444)
  AUTHORS   Smith CJ, Castanon O, Said K, Volf V, Khoshakhlagh P, Hornick A, 
            Ferreira R, Wu CT, Guell M, Garg S, Ng AHM, Myllykallio H, Church GM
  TITLE     Enabling large-scale genome editing at repetitive elements by 
            reducing DNA nicking.
  JOURNAL   Nucleic Acids Res. 2020 May 21;48(9):5183-5195. doi: 
            10.1093/nar/gkaa239.
  PUBMED    32315033
REFERENCE   2  (bases 1 to 9444)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2020-05-21- 21"; volume: "48"; issue: 
            "9"; pages: "5183-5195"
COMMENT     Sequence Label: dCBE4-gam
FEATURES             Location/Qualifiers
     source          1..9444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      652..661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1228..1911
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             join(2008..5406,5407..5409,5410..6006,6007,6008..6010,
                     6011..6012,6013..6015,6016..6108)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(2008..2031,2032..2034,2035..4521,4522..4524,
                     4525..6108)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6139..6387
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6418..6666
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6679..6699
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6708..6725
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6748..6765)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6754..6978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7049..7065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7049..7065)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7062..7084)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7073..7089
                     /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(7097..7103,7104..7121,7122..7127))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7142..7163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7280..7297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7451..8039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7531..7550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8210..9001,9002..9070))
                     /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     8833..8852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9071..9175)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9254..9273)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 gctaaaccag caaaacgtat caagagtgcc gcagcggctt atgtgccaca aaaccgcgat
      721 gcggtgatta ccgatattaa acgcatcggg gatttacagc gcgaagcatc acgtctggaa
      781 acggaaatga atgatgccat cgcggaaatt acggagaaat ttgcggcccg gattgcaccg
      841 attaaaaccg atattgaaac cctttcaaaa ggcgttcagg gatggtgtga agcgaaccgc
      901 gacgaactga cgaacggcgg caaagtgaag acggcgaatc ttgtcaccgg tgatgtatcg
      961 tggcgggtcc gtccaccatc agtaagtatt cgtggtatgg atgcagtgat ggaaacgctg
     1021 gagcgtcttg gcctgcaacg ctttattcgc acgaagcagg aaatcaacaa ggaagcgatt
     1081 ttactggaac cgaaagcggt cgcaggcgtt gccggaatta cagttaaatc aggcattgag
     1141 gatttttcta ttattccatt tgaacaggaa gccggtatta gcggcagcga gactcccggg
     1201 acctcagagt ccgctacacc cgaaagtagc tcagagactg gcccagtggc tgtggacccc
     1261 acattgagac ggcggatcga gccccatgag tttgaggtat tcttcgatcc gagagagctc
     1321 cgcaaggaga cctgcctgct ttacgaaatt aattgggggg gccggcactc catttggcga
     1381 catacatcac agaacactaa caagcacgtc gaagtcaact tcatcgagaa gttcacgaca
     1441 gaaagatatt tctgtccgaa cacaaggtgc agcattacct ggtttctcag ctggagccca
     1501 tgcggcgaat gtagtagggc catcactgaa ttcctgtcaa ggtatcccca cgtcactctg
     1561 tttatttaca tcgcaaggct gtaccaccac gctgaccccc gcaatcgaca aggcctgcgg
     1621 gatttgatct cttcaggtgt gactatccaa attatgactg agcaggagtc aggatactgc
     1681 tggagaaact ttgtgaatta tagcccgagt aatgaagccc actggcctag gtatccccat
     1741 ctgtgggtac gactgtacgt tcttgaactg tactgcatca tactgggcct gcctccttgt
     1801 ctcaacattc tgagaaggaa gcagccacag ctgacattct ttaccatcgc tcttcagtct
     1861 tgtcattacc agcgactgcc cccacacatt ctctgggcca ccgggttgaa atctggtggt
     1921 tcttctggtg gttctagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa
     1981 agttctggtg gttcttctgg tggttctgat aaaaagtatt ctattggttt agccatcggc
     2041 actaattccg ttggatgggc tgtcataacc gatgaataca aagtaccttc aaagaaattt
     2101 aaggtgttgg ggaacacaga ccgtcattcg attaaaaaga atcttatcgg tgccctccta
     2161 ttcgatagtg gcgaaacggc agaggcgact cgcctgaaac gaaccgctcg gagaaggtat
     2221 acacgtcgca agaaccgaat atgttactta caagaaattt ttagcaatga gatggccaaa
     2281 gttgacgatt ctttctttca ccgtttggaa gagtccttcc ttgtcgaaga ggacaagaaa
     2341 catgaacggc accccatctt tggaaacata gtagatgagg tggcatatca tgaaaagtac
     2401 ccaacgattt atcacctcag aaaaaagcta gttgactcaa ctgataaagc ggacctgagg
     2461 ttaatctact tggctcttgc ccatatgata aagttccgtg ggcactttct cattgagggt
     2521 gatctaaatc cggacaactc ggatgtcgac aaactgttca tccagttagt acaaacctat
     2581 aatcagttgt ttgaagagaa ccctataaat gcaagtggcg tggatgcgaa ggctattctt
     2641 agcgcccgcc tctctaaatc ccgacggcta gaaaacctga tcgcacaatt acccggagag
     2701 aagaaaaatg ggttgttcgg taaccttata gcgctctcac taggcctgac accaaatttt
     2761 aagtcgaact tcgacttagc tgaagatgcc aaattgcagc ttagtaagga cacgtacgat
     2821 gacgatctcg acaatctact ggcacaaatt ggagatcagt atgcggactt atttttggct
     2881 gccaaaaacc ttagcgatgc aatcctccta tctgacatac tgagagttaa tactgagatt
     2941 accaaggcgc cgttatccgc ttcaatgatc aaaaggtacg atgaacatca ccaagacttg
     3001 acacttctca aggccctagt ccgtcagcaa ctgcctgaga aatataagga aatattcttt
     3061 gatcagtcga aaaacgggta cgcaggttat attgacggcg gagcgagtca agaggaattc
     3121 tacaagttta tcaaacccat attagagaag atggatggga cggaagagtt gcttgtaaaa
     3181 ctcaatcgcg aagatctact gcgaaagcag cggactttcg acaacggtag cattccacat
     3241 caaatccact taggcgaatt gcatgctata cttagaaggc aggaggattt ttatccgttc
     3301 ctcaaagaca atcgtgaaaa gattgagaaa atcctaacct ttcgcatacc ttactatgtg
     3361 ggacccctgg cccgagggaa ctctcggttc gcatggatga caagaaagtc cgaagaaacg
     3421 attactccat ggaattttga ggaagttgtc gataaaggtg cgtcagctca atcgttcatc
     3481 gagaggatga ccaactttga caagaattta ccgaacgaaa aagtattgcc taagcacagt
     3541 ttactttacg agtatttcac agtgtacaat gaactcacga aagttaagta tgtcactgag
     3601 ggcatgcgta aacccgcctt tctaagcgga gaacagaaga aagcaatagt agatctgtta
     3661 ttcaagacca accgcaaagt gacagttaag caattgaaag aggactactt taagaaaatt
     3721 gaatgcttcg attctgtcga gatctccggg gtagaagatc gatttaatgc gtcacttggt
     3781 acgtatcatg acctcctaaa gataattaaa gataaggact tcctggataa cgaagagaat
     3841 gaagatatct tagaagatat agtgttgact cttaccctct ttgaagatcg ggaaatgatt
     3901 gaggaaagac taaaaacata cgctcacctg ttcgacgata aggttatgaa acagttaaag
     3961 aggcgtcgct atacgggctg gggacgattg tcgcggaaac ttatcaacgg gataagagac
     4021 aagcaaagtg gtaaaactat tctcgatttt ctaaagagcg acggcttcgc caataggaac
     4081 tttatgcagc tgatccatga tgactcttta accttcaaag aggatataca aaaggcacag
     4141 gtttccggac aaggggactc attgcacgaa catattgcga atcttgctgg ttcgccagcc
     4201 atcaaaaagg gcatactcca gacagtcaaa gtagtggatg agctagttaa ggtcatggga
     4261 cgtcacaaac cggaaaacat tgtaatcgag atggcacgcg aaaatcaaac gactcagaag
     4321 gggcaaaaaa acagtcgaga gcggatgaag agaatagaag agggtattaa agaactgggc
     4381 agccagatct taaaggagca tcctgtggaa aatacccaat tgcagaacga gaaactttac
     4441 ctctattacc tacaaaatgg aagggacatg tatgttgatc aggaactgga cataaaccgt
     4501 ttatctgatt acgacgtcga tgccattgta ccccaatcct ttttgaagga cgattcaatc
     4561 gacaataaag tgcttacacg ctcggataag aaccgaggga aaagtgacaa tgttccaagc
     4621 gaggaagtcg taaagaaaat gaagaactat tggcggcagc tcctaaatgc gaaactgata
     4681 acgcaaagaa agttcgataa cttaactaaa gctgagaggg gtggcttgtc tgaacttgac
     4741 aaggccggat ttattaaacg tcagctcgtg gaaacccgcc aaatcacaaa gcatgttgca
     4801 cagatactag attcccgaat gaatacgaaa tacgacgaga acgataagct gattcgggaa
     4861 gtcaaagtaa tcactttaaa gtcaaaattg gtgtcggact tcagaaagga ttttcaattc
     4921 tataaagtta gggagataaa taactaccac catgcgcacg acgcttatct taatgccgtc
     4981 gtagggaccg cactcattaa gaaatacccg aagctagaaa gtgagtttgt gtatggtgat
     5041 tacaaagttt atgacgtccg taagatgatc gcgaaaagcg aacaggagat aggcaaggct
     5101 acagccaaat acttctttta ttctaacatt atgaatttct ttaagacgga aatcactctg
     5161 gcaaacggag agatacgcaa acgaccttta attgaaacca atggggagac aggtgaaatc
     5221 gtatgggata agggccggga cttcgcgacg gtgagaaaag ttttgtccat gccccaagtc
     5281 aacatagtaa agaaaactga ggtgcagacc ggagggtttt caaaggaatc gattcttcca
     5341 aaaaggaata gtgataagct catcgctcgt aaaaaggact gggacccgaa aaagtacggt
     5401 ggcttcgata gccctacagt tgcctattct gtcctagtag tggcaaaagt tgagaaggga
     5461 aaatccaaga aactgaagtc agtcaaagaa ttattgggga taacgattat ggagcgctcg
     5521 tcttttgaaa agaaccccat cgacttcctt gaggcgaaag gttacaagga agtaaaaaag
     5581 gatctcataa ttaaactacc aaagtatagt ctgtttgagt tagaaaatgg ccgaaaacgg
     5641 atgttggcta gcgccggaga gcttcaaaag gggaacgaac tcgcactacc gtctaaatac
     5701 gtgaatttcc tgtatttagc gtcccattac gagaagttga aaggttcacc tgaagataac
     5761 gaacagaagc aactttttgt tgagcagcac aaacattatc tcgacgaaat catagagcaa
     5821 atttcggaat tcagtaagag agtcatccta gctgatgcca atctggacaa agtattaagc
     5881 gcatacaaca agcacaggga taaacccata cgtgagcagg cggaaaatat tatccatttg
     5941 tttactctta ccaacctcgg cgctccagcc gcattcaagt attttgacac aacgatagat
     6001 cgcaaacgat acacttctac caaggaggtg ctagacgcga cactgattca ccaatccatc
     6061 acgggattat atgaaactcg gatagatttg tcacagcttg ggggtgactc tggtggttct
     6121 ggaggatctg gtggttctac taatctgtca gatattattg aaaaggagac cggtaagcaa
     6181 ctggttatcc aggaatccat cctcatgctc ccagaggagg tggaagaagt cattgggaac
     6241 aagccggaaa gcgatatact cgtgcacacc gcctacgacg agagcaccga cgagaatgtc
     6301 atgcttctga ctagcgacgc ccctgaatac aagccttggg ctctggtcat acaggatagc
     6361 aacggtgaga acaagattaa gatgctctct ggtggttctg gaggatctgg tggttctact
     6421 aatctgtcag atattattga aaaggagacc ggtaagcaac tggttatcca ggaatccatc
     6481 ctcatgctcc cagaggaggt ggaagaagtc attgggaaca agccggaaag cgatatactc
     6541 gtgcacaccg cctacgacga gagcaccgac gagaatgtca tgcttctgac tagcgacgcc
     6601 cctgaataca agccttgggc tctggtcata caggatagca acggtgagaa caagattaag
     6661 atgctctctg gtggttctcc caagaagaag aggaaagtct aaccggtcat catcaccatc
     6721 accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     6781 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6841 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6901 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     6961 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga
     7021 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7081 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct
     7141 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7201 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7261 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7321 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7381 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7861 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7981 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8041 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8101 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8161 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8221 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8281 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8341 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8401 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8461 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8521 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8581 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8641 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8701 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8761 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8821 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8881 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8941 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9001 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9061 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9121 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9181 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag
     9241 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     9301 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     9361 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     9421 atgtacgggc cagatatacg cgtt
//
