LOCUS       sequence-209447-       17520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate point mutations using CRISPR/Cas9 system; encoded 
            cytidine deaminase fused to deactived Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17520)
  AUTHORS   Christopher Potter
  TITLE     Potter Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 17520)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pActin5c-CD-dCas9m4-UDI-attB
FEATURES             Location/Qualifiers
     source          1..17520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(675..693)
                     /label=pCasper-F
                     /note="Drosophila mini-white gene, reverse primer"
     CDS             join(1145..1216,1618..1891,1966..2620,2682..2997,3201..3332,
                     3403..4017)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     protein_bind    4792..4825
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4896..7406
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             9172..9858
                     /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(9907..9930,9931..9933,9934..12417,12418..12423,
                     12424..12489,12490..12492,12493..14007)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             14020..14268
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             14281..14301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          14394..14459
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             14589..14609
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    14881..15015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14918..14937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14972..14991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    15044..15113
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        complement(15332..15350)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(15371..15387)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15371..15387)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15384..15406)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15395..15411
                     /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(15419..15425,15426..15443,15444..15449))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15464..15485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15602..15619)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15773..16361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15853..15872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16532..17323,17324..17392))
                     /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     17155..17174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(17393..17497)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacgtaccg
      661 ggcccctagt atgtatgtaa gttaataaaa cccatttttg cggaaagtag ataaaaaaaa
      721 catttttttt ttttactgca ctggatatca ttgaacttat ctgatcagtt ttaaatttac
      781 ttcgatccaa gggtatttga tgtaccaggt tctttcgatt acctctcact caaaatgaca
      841 ttccactcaa agtcagcgct gtttgcctcc ttctctgtcc acagaaatat cgccgtctct
      901 ttcgccgctg cgtccgctat ctctttcgcc accgtttgta gcgttacgta gcgtcaatgt
      961 ccgccttcag ttgcattttg tcagcggttt cgtgacgaag ctccaagcgg tttacgccat
     1021 caattaaaca caaagtgctg tgccaaaact cctctcgctt cttatttttg tttgtttttt
     1081 gagtgattgg ggtggtgatt ggttttgggt gggtaagcag gggaaagtgt gaaaaatccc
     1141 ggcaatgggc caagaggatc aggagctatt aattcgcgga ggcagcaaac acccatctgc
     1201 cgagcatctg aacaatgtga gtagtacatg tgcatacatc ttaagttcac ttgatctata
     1261 ggaactgcga ttgcaacatc aaattgtctg cggcgtgaga actgcgaccc acaaaaatcc
     1321 caaaccgcaa ttgcacaaac aaatagtgac acgaaacaga ttattctggt agctgttctc
     1381 gctatataag acaatttttg agatcatatc atgatcaaga catctaaagg cattcatttt
     1441 cgactatatt cttttttaca aaaaatataa caaccagata ttttaagctg atcctagatg
     1501 cacaaaaaat aaataaaagt ataaacctac ttcgtaggat acttcggggt actttttgtt
     1561 cggggttaga tgagcataac gcttgtagtt gatatttgag atcccctatc attgcagggt
     1621 gacagcggag cggcttcgca gagctgcatt aaccagggct tcgggcaggc caaaaactac
     1681 ggcacgctcc ggccacccag tccgccggag gactccggtt cagggagcgg ccaactagcc
     1741 gagaacctca cctatgcctg gcacaatatg gacatctttg gggcggtcaa tcagccgggc
     1801 tccggatggc ggcagctggt caaccggaca cgcggactat tctgcaacga gcgacacata
     1861 ccggcgccca ggaaacattt gctcaagaac ggtgagtttc tattcgcagt cggctgatct
     1921 gtgtgaaatc ttaataaagg gtccaattac caatttgaaa ctcagtttgc ggcgtggcct
     1981 atccgggcga acttttggcc gtgatgggca gttccggtgc cggaaagacg accctgctga
     2041 atgcccttgc ctttcgatcg ccgcagggca tccaagtatc gccatccggg atgcgactgc
     2101 tcaatggcca acctgtggac gccaaggaga tgcaggccag gtgcgcctat gtccagcagg
     2161 atgacctctt tatcggctcc ctaacggcca gggaacacct gattttccaa gccatggtgc
     2221 ggatgccacg acatctgacc tatcggcagc gagtggcccg cgtggatcag gtgatccagg
     2281 agctttcgct cagcaaatgt cagcacacga tcatcggtgt gcccggcagg gtgaaaggtc
     2341 tgtccggcgg agaaaggaag cgtctggcat tcgcctccga ggctctaacc gatccgccgc
     2401 ttctgatctg cgatgagccc acctccggac tggactcctt taccgcccac agcgtcgtcc
     2461 aggtgctgaa gaagctgtcg cagaagggca agaccgtcat cctgaccatt catcagccgt
     2521 cttccgagct gtttgagctc tttgacaaga tccttctgat ggccgagggc agggtagctt
     2581 tcttgggcac tcccagcgaa gccgtcgact tcttttccta gtgagttcga tgtgtttatt
     2641 aagggtatct agtattacat aacatctcaa ctcctatcca gcgtgggtgc ccagtgtcct
     2701 accaactaca atccggcgga cttttacgta caggtgttgg ccgttgtgcc cggacgggag
     2761 atcgagtccc gtgatcggat cgccaagata tgcgacaatt ttgccattag caaagtagcc
     2821 cgggatatgg agcagttgtt ggccaccaaa aatctggaga agccactgga gcagccggag
     2881 aatgggtaca cctacaaggc cacctggttc atgcagttcc gggcggtcct gtggcgatcc
     2941 tggctgtcgg tgctcaagga accactcctc gtaaaagtgc gacttattca gacaacggtg
     3001 agtggttcca gtggaaacaa atgatataac gcttacaatt cttggaaaca aattcgctag
     3061 attttagata gaattgcctg attccacacc cttcttagtt tttttcaatg agatgtatag
     3121 tttatagttt tgcagaagat aaataaattt catttaactc gcgaatatta atgagatgcg
     3181 agtaacattt taatttgcag atggttgcca tcttgattgg cctcatcttt ttgggccaac
     3241 aactcacgca agtgggtgtg atgaatatca acggagccat cttcctcttc ctgaccaaca
     3301 tgacctttca aaacgtcttt gccacgataa atgtaagtca tgtttagaat acatttgcat
     3361 ttcaataatt tactaacttt ctaatgaatc gattcgattt aggtgttcac ctcagagctg
     3421 ccagttttta tgagggaggc ccgaagtcga ctttatcgct gtgacacata ctttctgggc
     3481 aaaacgattg ccgaattgcc gctttttctc acagtgccac tggtcttcac ggcgattgcc
     3541 tatccgatga tcggactgcg ggccggagtg ctgcacttct tcaactgcct ggcgctggtc
     3601 actctggtgg ccaatgtgtc aacgtccttc ggatatctaa tatcctgcgc cagctcctcg
     3661 acctcgatgg cgctgtctgt gggtccgccg gttatcatac cattcctgct ctttggcggc
     3721 ttcttcttga actcgggctc ggtgccagta tacctcaaat ggttgtcgta cctctcatgg
     3781 ttccgttacg ccaacgaggg tctgctgatt aaccaatggg cggacgtgga gccgggcgaa
     3841 attagctgca catcgtcgaa caccacgtgc cccagttcgg gcaaggtcat cctggagacg
     3901 cttaacttct ccgccgccga tctgccgctg gactacgtgg gtctggccat tctcatcgtg
     3961 agcttccggg tgctcgcata tctggctcta agacttcggg cccgacgcaa ggagtagccg
     4021 acatatatcc gaaataactg cttgtttttt ttttttacca ttattaccat cgtgtttact
     4081 gtttattgcc ccctcaaaaa gctaatgtaa ttatatttgt gccaataaaa acaagatatg
     4141 acctatagaa tacaagtatt tccccttcga acatccccac aagtagactt tggatttgtc
     4201 ttctaaccaa aagacttaca cacctgcata ccttacatca aaaactcgtt tatcgctaca
     4261 taaaacaccg ggatatattt tttatataca tacttttcaa atcgcgcgcc ctcttcataa
     4321 ttcacctcca ccacaccacg tttcgtagtt gctctttcgc tgtctcccac ccgctctccg
     4381 caacacattc accttttgtt cgacgacctt ggagcgactg tcgttagttc cgcgcgattc
     4441 ggttcgctca aatggttccg agtggttcat ttcgtctcaa tagaaattag taataaatat
     4501 ttgtatgtac aatttatttg ctccaatata tttgtatata tttccctcac agctatattt
     4561 attctaattt aatattatga ctttttaagg taattttttg tgacctgttc ggagtgatta
     4621 gcgttacaat ttgaactgaa agtgacatcc agtgtttgtt ccttgtgtag atgcatctca
     4681 aaaaaatggt gggcataata gtgttgttta tatatatcaa aaataacaac tataataata
     4741 agaatacatt taatttagaa aatgcttgga tttcactgga actaggctag cataacttcg
     4801 tataatgtat gctatacgaa gttatgctag cggatccaag cttgcggccg caattctata
     4861 ttctaaaaac acaaatgata cttctaaaaa aaaatcatga atggcatcaa ctctgaatca
     4921 aatctttgca gatgcaccta cttctcattt ccactgtcac atcatttttc cagatctcgc
     4981 tgcctgttat gtggcccaca aaccaagaca cgttttatgg ccattaaagc tggctgatcg
     5041 tcgccaaaca ccaaatacat aatgaatatg tacacattcg agaaagaagc gatcaaagaa
     5101 gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag aacgagctga tctagtatct
     5161 ctccacaatc caatgccaac tgaccaactg gccatattcg gagcaatttg aagccaattt
     5221 ccatcgcctg gcgatcgctc cattcttggc tatatgtttt tcaccgttac ccggggccat
     5281 tttcaaagac tcgtcggcaa gataagattg tgtcactcgc tgtctctctt catttgtcga
     5341 agaatgctga ggaatttcgc gatgacgtcg gcgagtattt tgaagaatga gaataatttg
     5401 tatttatacg aaaatcagtt agtggaattt tctacaaaaa catgttatct atagataatt
     5461 ttgttgcaaa atatgttgac tatgacaaag attgtatgta tataccttta atgtattctc
     5521 attttcttat gtatttataa tggcaatgat gatactgatg atattttaag atgatgccag
     5581 accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag tgcatgtgca attgttgttt
     5641 tttggaatat tcaattttcg gactgtccgc tttgatttca gtttcttggc ttattcaaaa
     5701 agcaaagtaa agccaaaaaa gcgagatggc aataccaaat gcggcaaaac ggtagtggaa
     5761 ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg gcgtggcggg gtctgtggct
     5821 gggcgcgacg tcaccgacgt tggagccact cctttgacca tgtgtgcgtg tgtgtattat
     5881 tcgtgtctcg ccactcgccg gttgtttttt tctttttatg ctgcgctctc tctagcgcca
     5941 tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt ccttttatgc gtcattttgg
     6001 ctcgaaatag gcaattattt aaacaaagat tagtcaacga aaacgctaaa ataaataagt
     6061 ctacaatatg gttacttatt gccatgtgtg tgcagccaac gatagcaaca aaagcaacaa
     6121 cacaggtggc tttccctctt tcactttttg tttgcaagcc gcgtgcgagc aagacggcac
     6181 gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag ttttggcgaa aaacatcaag
     6241 gcgcctgata cgaatgcatt tgcaataaca attgcgatat ttaatattgt ttatgaagct
     6301 gtttgacttc aaaacacaca aaaaaaaaaa ataaaacaaa ttatttgaaa gagaattagg
     6361 aatcggacgc ttatcgttag ggtaacaaca agaaatgctt actgagtcac agcctctgga
     6421 aaactgccgc aagccagaga gagagagaaa aagagggaga gcagcttaga ccgcatgtgc
     6481 ttgtgtgtga ggcgtctctc tcttcgtctc tgttgcgcaa acgcatagac tgcactgaaa
     6541 aaatcgatta cctatttttt atgaatgaat atttgcacta ttactattca aaactattaa
     6601 gatagcaatc acattcaata gccaaatact ataccacctg agcgatgcaa cgaaatgatc
     6661 aatttgagca aaaatgctgc atatttagga cggcatcatt atagaaatgc ttcttgctgt
     6721 gtacttttct ctcgtctggc agctgtttcg ccgttattgt taaaaccggc ttaagttagg
     6781 tgtgttttct acgactagtg aatgccctac tagaagatgt gtgttgcaca aaatgtccct
     6841 ggaataacca atttgaagtg cagatagcag taaacgtaag ctaatatgaa tattatttaa
     6901 ctgtaatgtt ttaatatcgc tggacattac taataaaccc actataaaca catgtacata
     6961 tgtatgtttt ggcatacaat gagtagttgg ggaaaaaatg tgtaaaagca ccgtgaccat
     7021 cacagcataa agataaccag ctgaagtatc gaatatgagt aacccccaaa ttgaatcaca
     7081 tgccgcaact gataggaccc atggaagtac actcttcatg gcgatataca agacacacac
     7141 aagcacgaac acccagttgc ggaggaaatt ctccgtaaat gaaaacccaa tcggcgaaca
     7201 attcataccc atatatggta aaagttttga acgcgacttg agagcggaga gcattgcggc
     7261 tgataaggtt ttagcgctaa gcgggcttta taaaacgggc tgcgggacca gttttcatat
     7321 cactaccgtt tgagttcttg tgctgtgtgg atactcctcc cgacacaaag ccgctccatc
     7381 agccagcagt cgtctaatcc agagacacca aaccgaaaga cttaatttat atttatttaa
     7441 ttaattttaa taaaacacac caaatgtaag tagctttccc cttcccaaca acaaaacacc
     7501 atcgaaccac tcccaccaag aaaaagcaat aatcgagaaa agccgcggaa aatgtgtgat
     7561 tttttttgta aacaaaacat ttttttatgt gccagtgctg aaagtgatca aaaaatacta
     7621 gccacgagct aaagagttat tgtattgacc aaaactccaa aaatacccaa gtttggccct
     7681 aaattgtcaa tcaaaatacc aataggtcga aagacatcaa aattaacaaa accagggttt
     7741 caaataccat aactcaagaa tcaggattac aactgcagat ttcaggatat atacatacaa
     7801 attatacgaa attataaaaa ccaaagcaat tcaatagccc caactcaaat gttaggatct
     7861 aatatagtgt ttaaagccaa gctcgctgat gtgggcgtgt cacgatttca cccaaagata
     7921 tgccaaatta cgaattgcaa atcaattcgc caacactttc tttttttccc acgccctaaa
     7981 acaccagatc atccataaat gtacatacat acagtatatg catattataa tctgtaaaac
     8041 tagatcaggt tcttgaaaat agtgacgtag gcagccgttt tggctgaagc agaaattttt
     8101 gccggttttt caaagttgta gttgcaaaaa tggagaaaac cttcgagcat tcgttcatat
     8161 acacacactc acgcgcaaaa taacgagaga gagtgtatgt gtgtgtgaga gagcgaaagc
     8221 cagacgacgg tttgcttttc gcctcgaaac atgaccatat atggtcacaa aacttggccg
     8281 ccgcaattca acacaccagc gctctccttc gcacccatag cgaccatggc gcggagcgag
     8341 cgagatggcg agagcgagcg acgcctatgg cgacgtcgac gcaggcagcg attgaaaaac
     8401 gcagttaact ggcattcaac attcaccagc cactttcagt cggtttattc cagtcattcc
     8461 tttcaaaccg tgcggtcgct tagctcagcc tcgccacttg cgtttacagt agttttcacg
     8521 ccttgaattt gttaaatcga acaaaaaggt aaagtttaac tagctttgaa aagtttcgtg
     8581 gctcttaatt gttaaatttt ctagagtgcg tttagtgttt tttttttttt ttattttgta
     8641 atgttaattt cgggttccaa ttcgagtttt aggcagccgc cattttaagg gcgcatacac
     8701 acaggcaact gtgctctctt tgcggctttc ttttgcaccg gcattcgtta agtgtcgtct
     8761 agaagcttct cccctccctt ttcggcatat tcgtattgtg gttttaattt ttcggggcgg
     8821 ggccttctat tttgtaactg ttcttttaat ttcttattac aattcgatcg caagtgaaaa
     8881 tcagttttca atcggaaaag tattttttta tgaaattttt ttttgtccaa gattaaaatt
     8941 ttgtactaaa aaaacgtaca ttgcattgag tgatttttaa ttgtacacga aaaacaagtt
     9001 agtttgttat gacaattgta ctttggtaga ccagcgcagt ccaaggaaac cacgcaaatt
     9061 ctcagttttt tttttgccat ttctacatta ccaaataagg taaccaaaaa ctaatgggaa
     9121 atccgcattc tttccattgc agcttacagg atcgatcccc gggaattcac catgagctca
     9181 gagactggcc cagtggctgt ggaccccaca ttgagacggc ggatcgagcc ccatgagttt
     9241 gaggtattct tcgatccgag agagctccgc aaggagacct gcctgcttta cgaaattaat
     9301 tgggggggcc ggcactccat ttggcgacat acatcacaga acactaacaa gcacgtcgaa
     9361 gtcaacttca tcgagaagtt cacgacagaa agatatttct gtccgaacac aaggtgcagc
     9421 attacctggt ttctcagctg gagcccatgc ggcgaatgta gtagggccat cactgaattc
     9481 ctgtcaaggt atccccacgt cactctgttt atttacatcg caaggctgta ccaccacgct
     9541 gacccccgca atcgacaagg cctgcgggat ttgatctctt caggtgtgac tatccaaatt
     9601 atgactgagc aggagtcagg atactgctgg agaaactttg tgaattatag cccgagtaat
     9661 gaagcccact ggcctaggta tccccatctg tgggtacgac tgtacgttct tgaactgtac
     9721 tgcatcatac tgggcctgcc tccttgtctc aacattctga gaaggaagca gccacagctg
     9781 acattcttta ccatcgctct tcagtcttgt cattaccagc gactgccccc acacattctc
     9841 tgggccaccg ggttgaaaag cggcagcgag actcccggga cctcagagtc cgccacaccc
     9901 gaaagtgata aaaagtattc tattggttta gccatcggca ctaattccgt tggatgggct
     9961 gtcataaccg atgaatacaa agtaccttca aagaaattta aggtgttggg gaacacagac
    10021 cgtcattcga ttaaaaagaa tcttatcggt gccctcctat tcgatagtgg cgaaacggca
    10081 gaggcgactc gcctgaaacg aaccgctcgg agaaggtata cacgtcgcaa gaaccgaata
    10141 tgttacttac aagaaatttt tagcaatgag atggccaaag ttgacgattc tttctttcac
    10201 cgtttggaag agtccttcct tgtcgaagag gacaagaaac atgaacggca ccccatcttt
    10261 ggaaacatag tagatgaggt ggcatatcat gaaaagtacc caacgattta tcacctcaga
    10321 aaaaagctag ttgactcaac tgataaagcg gacctgaggt taatctactt ggctcttgcc
    10381 catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc
    10441 gatgtcgata aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac
    10501 ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc
    10561 cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt
    10621 aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc
    10681 gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg
    10741 gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc
    10801 attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct
    10861 agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc
    10921 agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac
    10981 gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc
    11041 ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg
    11101 cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg
    11161 cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag
    11221 attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat
    11281 tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag
    11341 gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat
    11401 aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca
    11461 gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc
    11521 ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt
    11581 accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa
    11641 atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa
    11701 atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt
    11761 gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac
    11821 gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg
    11881 gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc
    11941 ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat
    12001 gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt
    12061 cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag
    12121 accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc
    12181 gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa
    12241 aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac
    12301 ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc
    12361 agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggct
    12421 gctatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga
    12481 tccgataaag ctagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg
    12541 aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat
    12601 ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg
    12661 cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg
    12721 aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag
    12781 tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac
    12841 aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa
    12901 aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg
    12961 aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac
    13021 agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag
    13081 cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat
    13141 ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa
    13201 gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg
    13261 atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc
    13321 gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc
    13381 gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc
    13441 gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc
    13501 aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag
    13561 ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc
    13621 agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg
    13681 gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt cagtaagaga
    13741 gtcatcctag ctgatgccaa tctggacaaa gtattaagcg catacaacaa gcacagggat
    13801 aaacccatac gtgagcaggc ggaaaatatt atccatttgt ttactcttac caacctcggc
    13861 gctccagccg cattcaagta ttttgacaca acgatagatc gcaaacgata cacttctacc
    13921 aaggaggtgc tagacgcgac actgattcac caatccatca cgggattata tgaaactcgg
    13981 atagatttgt cacagcttgg gggtgactct ggtggttcta ctaatctgtc agatattatt
    14041 gaaaaggaga ccggtaagca actggttatc caggaatcca tcctcatgct cccagaggag
    14101 gtggaagaag tcattgggaa caagccggaa agcgatatac tcgtgcacac cgcctacgac
    14161 gagagcaccg acgagaatgt catgcttctg actagcgacg cccctgaata caagccttgg
    14221 gctctggtca tacaggatag caacggtgag aacaagatta agatgctctc tggtggttct
    14281 cccaagaaga agaggaaagt ctaaggtacc tctagaggat ctttgtgaag gaaccttact
    14341 tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata
    14401 taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga
    14461 ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc
    14521 tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt
    14581 ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct tcagaattgc
    14641 taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca
    14701 ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat ttgatgtata
    14761 gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta
    14821 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
    14881 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    14941 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    15001 tatcatgtct ggatccacta gtgtcgacga tgtaggtcac ggtctcgaag ccgcggtgcg
    15061 ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc cacctcaccc atctggtcca
    15121 tcatgatgaa cgggtcgagg tggcggtagt tgatcccggc gaacgcgcgg cgcaccggga
    15181 agccctcgcc ctcgaaaccg ctgggcgcgg tggtcacggt gagcacggga cgtgcgacgg
    15241 cgtcggcggg tgcggatacg cggggcagcg tcagcgggtt ctcgacggtc acggcgggca
    15301 tgtcgacact agttctagcc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
    15361 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    15421 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    15481 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    15541 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    15601 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    15661 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    15721 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    15781 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    15841 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    15901 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    15961 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    16021 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    16081 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    16141 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    16201 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    16261 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    16321 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    16381 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    16441 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    16501 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    16561 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    16621 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    16681 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    16741 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    16801 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    16861 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    16921 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    16981 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    17041 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    17101 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    17161 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    17221 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    17281 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    17341 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    17401 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    17461 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
