LOCUS       sequence-380006-       10289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TadDE in mammalian cells with BlastR selection, Tol2 
            integration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10289)
  AUTHORS   Neugebauer ME, Hsu A, Arbab M, Krasnow NA, McElroy AN, Pandey S, 
            Doman JL, Huang TP, Raguram A, Banskota S, Newby GA, Tolar J, Osborn
            MJ, Liu DR
  TITLE     Evolution of an adenine base editor into a small, efficient cytosine
            base editor with low off-target activity.
  JOURNAL   Nat Biotechnol. 2022 Nov 10. pii: 10.1038/s41587-022-01533-6. doi: 
            10.1038/s41587-022-01533-6.
  PUBMED    36357719
REFERENCE   2  (bases 1 to 10289)
  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 
            Biotechnol. 2022 Nov 10. pii: 10.1038/s41587-022-01533-6. doi: 
            10.1038/s41587-022-01533-6."
COMMENT     Sequence Label: p2T-cmv-SpCas9-TadDE-BlastR
FEATURES             Location/Qualifiers
     source          1..10289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     22..41
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        22..40
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             90..110
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(708..731,732..734,735..4808)
                     /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             708..4808
                     /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="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             4839..5087
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5118..5366
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5409..5429
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5438..5455
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5478..5495)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5484..5708
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5779..5795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5779..5795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5792..5814)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5803..5819
                     /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(5827..5833,5834..5851,5852..5857))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5872..5893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    6010..6131
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6021..6040)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6075..6094
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6289..6687)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(6706..6753)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6801..7130)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6815..6950
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6869..6888)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7115..7135
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     misc_feature    join(7254..7441,7442..7453)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     rep_origin      complement(7514..8102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7594..7613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8273..9064,9065..9133))
                     /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     8896..8915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9134..9238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9317..9336)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    join(9528..9539,9540..9677)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     enhancer        9686..10065
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10066..10269
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10220..10240
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ccgctagaga tccgcggccg ctaatacgac tcactatagg gagagccgcc accatgaaac
       61 ggacagccga cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc agttctgagg
      121 tggagttttc ccacgagtac tggatgagac atgccctgac cctggccaag agggcacggg
      181 atgagggaga ggcgcctgtg ggagccgtgc tggtgctgaa caatagagtg atcggcgagg
      241 gctggaacag acgtatcggc ctgcacgacc caacagccca tgccgaaatt atggccctga
      301 gacagggcgg cctggtcatg cagaactcca gactgattga cgccaccctg tacgtgacat
      361 tcgagccttg cgtgatgtgc gccggcgcca tgatcaactc taggatcggc cgcgtggtgt
      421 ttggcgtgag gaactcaaaa agaggcgccg caggctccct gatgaacgtg ctgaactacc
      481 ccggcatgaa tcaccgcgtc gaaattaccg agggaatcct ggcagatgaa tgtgccgccc
      541 tgctgtgcga tttctatcgg atgcctagac aggtgttcaa tgctcagaag aaggcccaga
      601 gctccatcaa ctctggcgga tctagcggag gatcctctgg cagcgagaca ccaggaacaa
      661 gcgagtcagc aacaccagag agcagtggcg gcagcagcgg cggcagcgac aagaagtaca
      721 gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
      781 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
      841 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
      901 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
      961 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     1021 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     1081 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     1141 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     1201 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     1261 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     1321 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     1381 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     1441 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     1501 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     1561 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     1621 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     1681 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     1741 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     1801 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     1861 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     1921 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     1981 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     2041 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     2101 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     2161 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     2221 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     2281 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     2341 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     2401 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     2461 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     2521 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     2581 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     2641 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     2701 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     2761 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     2821 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     2881 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     2941 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     3001 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     3061 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     3121 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     3181 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     3241 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     3301 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     3361 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     3421 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     3481 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     3541 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     3601 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     3661 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     3721 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     3781 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     3841 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     3901 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     3961 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     4021 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     4081 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     4141 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     4201 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     4261 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     4321 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     4381 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     4441 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     4501 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     4561 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     4621 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     4681 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     4741 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     4801 gaggtgacag cggcgggagc ggcgggagcg gggggagcac taatctgagc gacatcattg
     4861 agaaggagac tgggaaacag ctggtcattc aggagtccat cctgatgctg cctgaggagg
     4921 tggaggaagt gatcggcaac aagccagagt ctgacatcct ggtgcacacc gcctacgacg
     4981 agtccacaga tgagaatgtg atgctgctga cctctgacgc ccccgagtat aagccttggg
     5041 ccctggtcat ccaggattct aacggcgaga ataagatcaa gatgctgagc ggaggatccg
     5101 gaggatctgg aggcagcacc aacctgtctg acatcatcga gaaggagaca ggcaagcagc
     5161 tggtcatcca ggagagcatc ctgatgctgc ccgaagaagt cgaagaagtg atcggaaaca
     5221 agcctgagag cgatatcctg gtccataccg cctacgacga gagtaccgac gaaaatgtga
     5281 tgctgctgac atccgacgcc ccagagtata agccctgggc tctggtcatc caggattcca
     5341 acggagagaa caaaatcaaa atgctgtctg gcggctcaaa aagaaccgcc gacggcagcg
     5401 aattcgagcc caagaagaag aggaaagtct aaccggtcat catcaccatc accattgagt
     5461 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     5521 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     5581 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     5641 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     5701 ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga cctctagcta
     5761 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5821 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct aatgagtgag
     5881 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5941 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcta
     6001 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     6061 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     6121 ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     6181 cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa
     6241 tcgaaatctc gtagcacgtg tcagtcctgc tcctcggcca cgaagtgctt agccctccca
     6301 cacataacca gagggcagca attcacgaat cccaactgcc gtcggctgtc catcactgtc
     6361 cttcactatg gctttgatcc caggatgcag atcgagaagc acctgtcggc accgtccgca
     6421 ggggctcaag atgcccctgt tctcatttcc gatcgcgacg atacaagtca ggttgccagc
     6481 tgccgcagca gcagcagtgc ccagcaccac gagttctgca caaggtcccc cagtaaaatg
     6541 atatacattg acaccagtga agatgcggcc gtcgctagag agagctgcgc tggcgacgct
     6601 gtagtcttca gagatgggga tgctgttgat tgtagccgtt gctctttcaa tgagggtgga
     6661 ttcttcttga gacaaaggct tggccatggt ttagttcctc accttgtcgt attatactat
     6721 gccgatatac tatgccgatg attaattgtc aacacgtgct gatcagatcc gaaaatggat
     6781 atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     6841 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
     6901 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     6961 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     7021 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
     7081 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
     7141 attcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtaagtgat
     7261 ctccaaaaaa taagtacttt ttgactgtaa ataaaattgt aaggagtaaa aagtactttt
     7321 ttttctaaaa aaatgtaatt aagtaaaagt aaaagtattg atttttaatt gtactcaagt
     7381 aaagtaaaaa tccccaaaaa taatacttaa gtacagtaat caagtaaaat tactcaagta
     7441 ctttacacct ctgcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8041 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8101 aagaagatcc tttgatcttt tctacggggt ctgacactca gtggaacgaa aactcacgtt
     8161 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8221 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8281 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8341 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8401 caatgatacc gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8461 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8521 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8581 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8641 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8701 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8761 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8821 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8881 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8941 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9001 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9061 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9121 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9181 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9241 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
     9301 tagggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     9361 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     9421 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     9481 gcgatgtacg ggccagatat acgcgggtac cggcatatgg ttcttgacag aggtgtaaaa
     9541 agtactcaaa aattttactc aagtgaaagt acaagtactt agggaaaatt ttactcaatt
     9601 aaaagtaaaa gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt actccattaa
     9661 aattgtactt gagtattaac gcgttgacat tgattattga ctagttatta atagtaatca
     9721 attacggggt cattagttta tagcccatat atggagttcc gcgttacata acttacggta
     9781 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     9841 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     9901 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     9961 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
    10021 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
    10081 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
    10141 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
    10201 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
    10261 agcagagctg gtttagtgaa ccgtcagat
//
