LOCUS       sequence-267697-        9114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of BE3 (Base editor generation three) with dual 
            C-terminal NLS in a single operon encoding P2A blasticidine 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9114)
  AUTHORS   Nguyen Tran MT, Mohd Khalid MKN, Wang Q, Walker JKR, Lidgerwood GE, 
            Dilworth KL, Lisowski L, Pebay A, Hewitt AW
  TITLE     Engineering domain-inlaid SaCas9 adenine base editors with reduced 
            RNA off-targets and increased on-target DNA editing.
  JOURNAL   Nat Commun. 2020 Sep 25;11(1):4871. doi: 10.1038/s41467-020-18715-y.
  PUBMED    32978399
REFERENCE   2  (bases 1 to 9114)
  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: "2020-09-25"; volume: "11(1):4871. doi"; pages: " 
            10.1038/s41467-020-18715-y"
COMMENT     Sequence Label: BE3-NLS-NLS-P2A-BLAST
FEATURES             Location/Qualifiers
     source          1..9114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        89..468
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        469..672
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     623..643
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     714..733
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        714..732
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             746..1432
                     /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="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(1481..4879,4880..4882,4883..5479,5480,5481..5483,
                     5484..5485,5486..5488,5489..5581)
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1481..1504,1505..1507,1508..5581)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5594..5842
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5855..5875
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5888..5908
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5921..5968
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5969..5992
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6002..6058
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6059..6457
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             6466..6483
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6506..6523)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6512..6736
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6807..6823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6807..6823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6820..6842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6831..6847
                     /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(6855..6861,6862..6879,6880..6885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6900..6921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7038..7055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7209..7797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7289..7308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7968..8759,8760..8828))
                     /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     8591..8610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8829..8933)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9012..9031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tacgctgctt
       61 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
      121 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      181 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      241 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      301 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      361 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      421 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      481 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      541 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      601 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      661 ataagcagag ctggtttagt gaaccgtcag atccgctaga gatccgcggc cgctaatacg
      721 actcactata gggagagccg ccaccatgag ctcagagact ggcccagtgg ctgtggaccc
      781 cacattgaga cggcggatcg agccccatga gtttgaggta ttcttcgatc cgagagagct
      841 ccgcaaggag acctgcctgc tttacgaaat taattggggg ggccggcact ccatttggcg
      901 acatacatca cagaacacta acaagcacgt cgaagtcaac ttcatcgaga agttcacgac
      961 agaaagatat ttctgtccga acacaaggtg cagcattacc tggtttctca gctggagccc
     1021 atgcggcgaa tgtagtaggg ccatcactga attcctgtca aggtatcccc acgtcactct
     1081 gtttatttac atcgcaaggc tgtaccacca cgctgacccc cgcaatcgac aaggcctgcg
     1141 ggatttgatc tcttcaggtg tgactatcca aattatgact gagcaggagt caggatactg
     1201 ctggagaaac tttgtgaatt atagcccgag taatgaagcc cactggccta ggtatcccca
     1261 tctgtgggta cgactgtacg ttcttgaact gtactgcatc atactgggcc tgcctccttg
     1321 tctcaacatt ctgagaagga agcagccaca gctgacattc tttaccatcg ctcttcagtc
     1381 ttgtcattac cagcgactgc ccccacacat tctctgggcc accgggttga aaagcggcag
     1441 cgagactccc gggacctcag agtccgccac acccgaaagt gataaaaagt attctattgg
     1501 tttagccatc ggcactaatt ccgttggatg ggctgtcata accgatgaat acaaagtacc
     1561 ttcaaagaaa tttaaggtgt tggggaacac agaccgtcat tcgattaaaa agaatcttat
     1621 cggtgccctc ctattcgata gtggcgaaac ggcagaggcg actcgcctga aacgaaccgc
     1681 tcggagaagg tatacacgtc gcaagaaccg aatatgttac ttacaagaaa tttttagcaa
     1741 tgagatggcc aaagttgacg attctttctt tcaccgtttg gaagagtcct tccttgtcga
     1801 agaggacaag aaacatgaac ggcaccccat ctttggaaac atagtagatg aggtggcata
     1861 tcatgaaaag tacccaacga tttatcacct cagaaaaaag ctagttgact caactgataa
     1921 agcggacctg aggttaatct acttggctct tgcccatatg ataaagttcc gtgggcactt
     1981 tctcattgag ggtgatctaa atccggacaa ctcggatgtc gacaaactgt tcatccagtt
     2041 agtacaaacc tataatcagt tgtttgaaga gaaccctata aatgcaagtg gcgtggatgc
     2101 gaaggctatt cttagcgccc gcctctctaa atcccgacgg ctagaaaacc tgatcgcaca
     2161 attacccgga gagaagaaaa atgggttgtt cggtaacctt atagcgctct cactaggcct
     2221 gacaccaaat tttaagtcga acttcgactt agctgaagat gccaaattgc agcttagtaa
     2281 ggacacgtac gatgacgatc tcgacaatct actggcacaa attggagatc agtatgcgga
     2341 cttatttttg gctgccaaaa accttagcga tgcaatcctc ctatctgaca tactgagagt
     2401 taatactgag attaccaagg cgccgttatc cgcttcaatg atcaaaaggt acgatgaaca
     2461 tcaccaagac ttgacacttc tcaaggccct agtccgtcag caactgcctg agaaatataa
     2521 ggaaatattc tttgatcagt cgaaaaacgg gtacgcaggt tatattgacg gcggagcgag
     2581 tcaagaggaa ttctacaagt ttatcaaacc catattagag aagatggatg ggacggaaga
     2641 gttgcttgta aaactcaatc gcgaagatct actgcgaaag cagcggactt tcgacaacgg
     2701 tagcattcca catcaaatcc acttaggcga attgcatgct atacttagaa ggcaggagga
     2761 tttttatccg ttcctcaaag acaatcgtga aaagattgag aaaatcctaa cctttcgcat
     2821 accttactat gtgggacccc tggcccgagg gaactctcgg ttcgcatgga tgacaagaaa
     2881 gtccgaagaa acgattactc catggaattt tgaggaagtt gtcgataaag gtgcgtcagc
     2941 tcaatcgttc atcgagagga tgaccaactt tgacaagaat ttaccgaacg aaaaagtatt
     3001 gcctaagcac agtttacttt acgagtattt cacagtgtac aatgaactca cgaaagttaa
     3061 gtatgtcact gagggcatgc gtaaacccgc ctttctaagc ggagaacaga agaaagcaat
     3121 agtagatctg ttattcaaga ccaaccgcaa agtgacagtt aagcaattga aagaggacta
     3181 ctttaagaaa attgaatgct tcgattctgt cgagatctcc ggggtagaag atcgatttaa
     3241 tgcgtcactt ggtacgtatc atgacctcct aaagataatt aaagataagg acttcctgga
     3301 taacgaagag aatgaagata tcttagaaga tatagtgttg actcttaccc tctttgaaga
     3361 tcgggaaatg attgaggaaa gactaaaaac atacgctcac ctgttcgacg ataaggttat
     3421 gaaacagtta aagaggcgtc gctatacggg ctggggacga ttgtcgcgga aacttatcaa
     3481 cgggataaga gacaagcaaa gtggtaaaac tattctcgat tttctaaaga gcgacggctt
     3541 cgccaatagg aactttatgc agctgatcca tgatgactct ttaaccttca aagaggatat
     3601 acaaaaggca caggtttccg gacaagggga ctcattgcac gaacatattg cgaatcttgc
     3661 tggttcgcca gccatcaaaa agggcatact ccagacagtc aaagtagtgg atgagctagt
     3721 taaggtcatg ggacgtcaca aaccggaaaa cattgtaatc gagatggcac gcgaaaatca
     3781 aacgactcag aaggggcaaa aaaacagtcg agagcggatg aagagaatag aagagggtat
     3841 taaagaactg ggcagccaga tcttaaagga gcatcctgtg gaaaataccc aattgcagaa
     3901 cgagaaactt tacctctatt acctacaaaa tggaagggac atgtatgttg atcaggaact
     3961 ggacataaac cgtttatctg attacgacgt cgatcacatt gtaccccaat cctttttgaa
     4021 ggacgattca atcgacaata aagtgcttac acgctcggat aagaaccgag ggaaaagtga
     4081 caatgttcca agcgaggaag tcgtaaagaa aatgaagaac tattggcggc agctcctaaa
     4141 tgcgaaactg ataacgcaaa gaaagttcga taacttaact aaagctgaga ggggtggctt
     4201 gtctgaactt gacaaggccg gatttattaa acgtcagctc gtggaaaccc gccaaatcac
     4261 aaagcatgtt gcacagatac tagattcccg aatgaatacg aaatacgacg agaacgataa
     4321 gctgattcgg gaagtcaaag taatcacttt aaagtcaaaa ttggtgtcgg acttcagaaa
     4381 ggattttcaa ttctataaag ttagggagat aaataactac caccatgcgc acgacgctta
     4441 tcttaatgcc gtcgtaggga ccgcactcat taagaaatac ccgaagctag aaagtgagtt
     4501 tgtgtatggt gattacaaag tttatgacgt ccgtaagatg atcgcgaaaa gcgaacagga
     4561 gataggcaag gctacagcca aatacttctt ttattctaac attatgaatt tctttaagac
     4621 ggaaatcact ctggcaaacg gagagatacg caaacgacct ttaattgaaa ccaatgggga
     4681 gacaggtgaa atcgtatggg ataagggccg ggacttcgcg acggtgagaa aagttttgtc
     4741 catgccccaa gtcaacatag taaagaaaac tgaggtgcag accggagggt tttcaaagga
     4801 atcgattctt ccaaaaagga atagtgataa gctcatcgct cgtaaaaagg actgggaccc
     4861 gaaaaagtac ggtggcttcg atagccctac agttgcctat tctgtcctag tagtggcaaa
     4921 agttgagaag ggaaaatcca agaaactgaa gtcagtcaaa gaattattgg ggataacgat
     4981 tatggagcgc tcgtcttttg aaaagaaccc catcgacttc cttgaggcga aaggttacaa
     5041 ggaagtaaaa aaggatctca taattaaact accaaagtat agtctgtttg agttagaaaa
     5101 tggccgaaaa cggatgttgg ctagcgccgg agagcttcaa aaggggaacg aactcgcact
     5161 accgtctaaa tacgtgaatt tcctgtattt agcgtcccat tacgagaagt tgaaaggttc
     5221 acctgaagat aacgaacaga agcaactttt tgttgagcag cacaaacatt atctcgacga
     5281 aatcatagag caaatttcgg aattcagtaa gagagtcatc ctagctgatg ccaatctgga
     5341 caaagtatta agcgcataca acaagcacag ggataaaccc atacgtgagc aggcggaaaa
     5401 tattatccat ttgtttactc ttaccaacct cggcgctcca gccgcattca agtattttga
     5461 cacaacgata gatcgcaaac gatacacttc taccaaggag gtgctagacg cgacactgat
     5521 tcaccaatcc atcacgggat tatatgaaac tcggatagat ttgtcacagc ttgggggtga
     5581 ctctggtggt tctactaatc tgtcagatat tattgaaaag gagaccggta agcaactggt
     5641 tatccaggaa tccatcctca tgctcccaga ggaggtggaa gaagtcattg ggaacaagcc
     5701 ggaaagcgat atactcgtgc acaccgccta cgacgagagc accgacgaga atgtcatgct
     5761 tctgactagc gacgcccctg aatacaagcc ttgggctctg gtcatacagg atagcaacgg
     5821 tgagaacaag attaagatgc tctctggtgg ttctcccaag aagaagagga aagtctctgg
     5881 tggttctccc aagaagaaga ggaaagtctc tggtggttct aagcgacctg ccgccacaaa
     5941 gaaggctgga caggctaaga agaagaaaga ttacaaagac gatgacgata agggatccgg
     6001 cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgat
     6061 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     6121 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     6181 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     6241 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     6301 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     6361 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     6421 tgaattgctg ccctctggtt atgtgtggga gggctaataa ccggtcatca tcaccatcac
     6481 cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     6541 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     6601 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     6661 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     6721 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc
     6781 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa
     6901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6961 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7021 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7621 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8881 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8941 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg
     9001 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     9061 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aagg
//
