LOCUS       sequence-308861-       10472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-to-T base editor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10472)
  AUTHORS   Arbab M, Shen MW, Mok B, Wilson C, Matuszek Z, Cassa CA, Liu DR
  TITLE     Determinants of Base Editing Outcomes from Target Library Analysis 
            and Machine Learning.
  JOURNAL   Cell. 2020 Jun 9. pii: S0092-8674(20)30632-2. doi: 
            10.1016/j.cell.2020.05.037.
  PUBMED    32533916
REFERENCE   2  (bases 1 to 10472)
  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: "Cell. 2020 
            Jun 9. pii: S0092-8674(20)30632-2. doi: 10.1016/j.cell.2020.05.037."
COMMENT     Sequence Label: p2T-CMV-evoAmax-BlastR
FEATURES             Location/Qualifiers
     source          1..10472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     48..67
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        48..66
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             116..136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(917..940,941..943,944..5017)
                     /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             917..5017
                     /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             5048..5296
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5327..5575
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5618..5638
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5647..5664
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5687..5704)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5693..5917
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5988..6004)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5988..6004)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6001..6023)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6012..6028
                     /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(6036..6042,6043..6060,6061..6066))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6081..6102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    6219..6340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6230..6249)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6284..6303
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6498..6896)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(6915..6962)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(7010..7339)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7024..7159
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7078..7097)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7324..7344
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     misc_feature    join(7463..7650,7651..7662)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     rep_origin      complement(7723..8311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7803..7822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8482..9273,9274..9342))
                     /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     9105..9124
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9343..9447)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9526..9545)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    join(9737..9748,9749..9886)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     enhancer        9895..10274
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(10275..10472,1..6)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10429..10449
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 agagctggtt tagtgaaccg tcagatccgc tagagatccg cggccgctaa tacgactcac
       61 tatagggaga gccgccacca tgaaacggac agccgacgga agcgagttcg agtcaccaaa
      121 gaagaagcgg aaagtcagtt caaagactgg gcctgtcgcc gtcgatccaa ccctgcgccg
      181 ccggattgaa cctcacgagt ttgaagtgtt ctttgacccc cgggagctga gaaaggagac
      241 atgcctgctg tacgagatca actggggagg caggcactcc atctggaggc acacctctca
      301 gaacacaaat aagcacgtgg aggtgaactt catcgagaag tttaccacag agcggtactt
      361 ctgccccaat accagatgta gcatcacatg gtttctgagc tggtcccctt gcggagagtg
      421 tagcagggcc atcaccgagt tcctgtccag atatccaaat gtgacactgt ttatctacat
      481 cgccaggctg tatcacctgg caaacccaag gaataggcag ggcctgcgcg atctgatcag
      541 ctccggcgtg accatccaga tcatgacaga gcaggagtcc ggctactgct ggcacaactt
      601 cgtgaattat tctcctagca acgagtccca ctggcctagg tacccacacc tgtgggtgcg
      661 cctgtacgtg ctggagctgt attgcatcat cctgggcctg cccccttgtc tgaatatcct
      721 gcggagaaag cagagccagc tgacctcctt tacaatcgcc ctgcagtctt gtcactatca
      781 gaggctgcca ccccacatcc tgtgggccac aggcctgaag tctggcggat ctagcggagg
      841 atcctctggc agcgagacac caggaacaag cgagtcagca acaccagaga gcagtggcgg
      901 cagcagcggc ggcagcgaca agaagtacag catcggcctg gccatcggca ccaactctgt
      961 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     1021 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
     1081 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     1141 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     1201 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     1261 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     1321 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     1381 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     1441 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     1501 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     1561 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     1621 cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     1681 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     1741 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     1801 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     1861 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     1921 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     1981 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     2041 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     2101 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     2161 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     2221 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     2281 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     2341 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     2401 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     2461 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     2521 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     2581 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     2641 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     2701 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     2761 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     2821 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     2881 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     2941 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     3001 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     3061 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     3121 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     3181 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     3241 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     3301 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     3361 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     3421 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     3481 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     3541 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     3601 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     3661 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     3721 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     3781 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     3841 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     3901 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     3961 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     4021 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     4081 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     4141 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     4201 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     4261 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     4321 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     4381 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     4441 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     4501 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     4561 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     4621 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     4681 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     4741 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     4801 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     4861 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     4921 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     4981 cgagacacgg atcgacctgt ctcagctggg aggtgacagc ggcgggagcg gcgggagcgg
     5041 ggggagcact aatctgagcg acatcattga gaaggagact gggaaacagc tggtcattca
     5101 ggagtccatc ctgatgctgc ctgaggaggt ggaggaagtg atcggcaaca agccagagtc
     5161 tgacatcctg gtgcacaccg cctacgacga gtccacagat gagaatgtga tgctgctgac
     5221 ctctgacgcc cccgagtata agccttgggc cctggtcatc caggattcta acggcgagaa
     5281 taagatcaag atgctgagcg gaggatccgg aggatctgga ggcagcacca acctgtctga
     5341 catcatcgag aaggagacag gcaagcagct ggtcatccag gagagcatcc tgatgctgcc
     5401 cgaagaagtc gaagaagtga tcggaaacaa gcctgagagc gatatcctgg tccataccgc
     5461 ctacgacgag agtaccgacg aaaatgtgat gctgctgaca tccgacgccc cagagtataa
     5521 gccctgggct ctggtcatcc aggattccaa cggagagaac aaaatcaaaa tgctgtctgg
     5581 cggctcaaaa agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcta
     5641 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     5701 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     5761 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     5821 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     5881 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     5941 tggggctcga taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     6001 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6061 tgtaaagcct agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6121 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6181 gggagaggcg gtttgcgtat tgggcgctac agacatgata agatacattg atgagtttgg
     6241 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6301 tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg aagaactcca
     6361 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     6421 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tagcacgtgt cagtcctgct
     6481 cctcggccac gaagtgctta gccctcccac acataaccag agggcagcaa ttcacgaatc
     6541 ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga
     6601 tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg
     6661 atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg
     6721 agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg
     6781 tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt
     6841 gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtt
     6901 tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga ttaattgtca
     6961 acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc
     7021 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct
     7081 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     7141 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     7201 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     7261 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     7321 cacaccctaa ctgacacaca ttccacagaa ttcactgact cgctgcgctc ggtcgttcgg
     7381 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7441 gataacgcag gaaagaacat gtaagtgatc tccaaaaaat aagtactttt tgactgtaaa
     7501 taaaattgta aggagtaaaa agtacttttt tttctaaaaa aatgtaatta agtaaaagta
     7561 aaagtattga tttttaattg tactcaagta aagtaaaaat ccccaaaaat aatacttaag
     7621 tacagtaatc aagtaaaatt actcaagtac tttacacctc tgcatgtgag caaaaggcca
     7681 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7741 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7801 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7861 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7921 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7981 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8041 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8101 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8161 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8221 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8281 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8341 tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8401 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8461 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8521 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8581 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgggacccac gctcaccggc
     8641 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8701 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8761 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8821 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8881 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8941 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9001 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9061 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9121 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9181 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9241 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9301 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9361 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9421 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     9481 cggatcggga gatcgatctc ccgatcccct aaggtcgact ctcagtacaa tctgctctga
     9541 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     9601 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     9661 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgggtacc
     9721 ggcatatggt tcttgacaga ggtgtaaaaa gtactcaaaa attttactca agtgaaagta
     9781 caagtactta gggaaaattt tactcaatta aaagtaaaag tatctggcta gaatcttact
     9841 tgagtaaaag taaaaaagta ctccattaaa attgtacttg agtattaacg cgttgacatt
     9901 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     9961 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
    10021 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
    10081 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
    10141 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
    10201 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
    10261 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
    10321 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
    10381 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
    10441 gtaggcgtgt acggtgggag gtctatataa gc
//
