LOCUS       sequence-412695-        9321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of evoCjCas9 CBE4max.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9321)
  AUTHORS   Schmidheini L, Mathis N, Marquart KF, Rothgangl T, Kissling L, Bock 
            D, Chanez C, Wang JP, Jinek M, Schwank G
  TITLE     Continuous directed evolution of a compact CjCas9 variant with broad
            PAM compatibility.
  JOURNAL   Nat Chem Biol. 2023 Sep 21. doi: 10.1038/s41589-023-01427-x.
  PUBMED    37735239
REFERENCE   2  (bases 1 to 9321)
  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 Chem 
            Biol. 2023 Sep 21. doi: 10.1038/s41589-023-01427-x."
COMMENT     Sequence Label: p2T-CMV-evoCjCas9-BE4max-BlastR
FEATURES             Location/Qualifiers
     source          1..9321
                     /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             137..820
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             3896..4144
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             4175..4423
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             4466..4486
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4496..4513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4536..4553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4542..4766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4837..4853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4837..4853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4850..4872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4861..4877
                     /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(4885..4891,4892..4909,4910..4915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4930..4951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    5068..5189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5079..5098)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5133..5152
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5347..5745)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(5764..5811)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5859..6188)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5873..6008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5927..5946)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6173..6193
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     misc_feature    join(6312..6499,6500..6511)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     rep_origin      complement(6572..7160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6652..6671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7331..8122,8123..8191))
                     /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     7954..7973
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8192..8296)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8375..8394)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    join(8586..8597,8598..8735)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     enhancer        8744..9123
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(9124..9321,1..6)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9278..9298
                     /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 aaagtctcct cagagactgg 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 atatccacac gtgacactgt ttatctacat
      481 cgccaggctg tatcaccacg cagacccaag gaataggcag ggcctgcgcg atctgatcag
      541 ctccggcgtg accatccaga tcatgacaga gcaggagtcc ggctactgct ggcggaactt
      601 cgtgaattat tctcctagca acgaggccca ctggcctagg tacccacacc tgtgggtgcg
      661 cctgtacgtg ctggagctgt attgcatcat cctgggcctg cccccttgtc tgaatatcct
      721 gcggagaaag cagccccagc tgaccttctt tacaatcgcc ctgcagtctt gtcactatca
      781 gaggctgcca ccccacatcc tgtgggccac aggcctgaag tctggaggat ctagcggagg
      841 atcctctggc agcgagacac caggaacaag cgagtcagca acaccagaga gcagtggcgg
      901 cagcagcggc gggtcagccc gcatcctggc ctttgccatc ggcatcagca gcatcggatg
      961 ggccttcagc gagaatgacg agctgaagga ctgcggtgtg agaatcttta caaaggtcga
     1021 gaaccccaag accggcgaaa gcctggcgct gccaagacgg ctggcccggt ctgctcggaa
     1081 gagatacgcc cggagaaaag ccagactgaa tcacctgaaa cacctgatcg ccaacgagtt
     1141 caaactgaac tacgaggact accagagctt cgatgagagc ttggctaagg cctacaaagg
     1201 cagcctgatc agcccctacg agctgagatt cagagctctg aatgagctcc tgagcaagca
     1261 ggacttcgct agagtgatcc tgcacatcgc caagcgaaga ggctacgacg acatcaagaa
     1321 ctctgatgac aaggagaagg gcgccattct gaaagccatc aagcagaacg aagagaaact
     1381 ggccaattac cagagcgtgg gcgagtacct gtacaaggag tacttccaga agttcaaaga
     1441 aaatagcaaa gagttcacca acgtgcggaa caagaaggag tcttatgaaa gatgtatcgc
     1501 ccagagcttc ctgaaagacg agctgaaact gatcttcaag aagcaaagag agtttggctt
     1561 cagcttcagc aaaaaatttg aagaagaggt gctgtctgtc gccttctaca agagggctct
     1621 gaaggacttc agccacctgg tgggcaattg cagctttttc acagacgaaa agcgggcccc
     1681 taagaacagc cctctggcct tcatgttcgt ggctctgacc agaatcatca acctgctgaa
     1741 caacctgaaa aataccgagg gcatcctcta taccaaggac gatctgaacg ccctgctgaa
     1801 tgaggtgctc aagaatggca ccctgaccta caagcaaaca aaaaaactgc tgggcctgtc
     1861 tgacgactac gagtttaaag gcgagaaggg cacctatttc atcgaattta agaagtacaa
     1921 ggaattcatc aaggcactgg gcgaacacaa cctgtctcag gacgacctga acgagatcgc
     1981 caaggacatc accctgatca aggatgagat caagctgaaa aaagctctgg ccaagtacga
     2041 cctgaatcaa aaccagatcg acagcttaag caagctggaa tttaaggatc acctgaacat
     2101 ctcctttaag gccctgaagc tggtgacccc actgatgctg gaaggaaaga agtacgacga
     2161 agcatgcaac gagcttaacc tgaaggttgc tatcaacgag gataagaagg atttcctgcc
     2221 tgcctttaac gagacatact acaaggatga ggtgaccaac cccgtggtgc tgagagctat
     2281 caaagagtac agaaaggtgc tgaacgccct gctgaagaag tacggcaagg tccacaagat
     2341 caatattgag ctggcccggg aggttggaaa gaaccactct cagagagcaa agatcgagaa
     2401 ggagcaaaac gagaactaca aagcgaagaa ggacgccgaa ctggagtgcg agaagcttgg
     2461 cctgaagatc aactctaaga atatcctgaa actcagactt ttcaaagaac agaaggaatt
     2521 ctgtgcctac agcggcgaga agatcaaaat ttctgacctg caggatgaaa agatgctgga
     2581 gatcgaccac atctaccctt actccagaag cttcgacgac agctatatga acaaagtgct
     2641 ggtgttcaca aagcagaacc aggagaagct gaatcagacc cctttcgagg ccttcggcaa
     2701 tgactccgcc aagtggcaga aaatcgaggt gctggccaaa aacctgccaa ccaagaaaca
     2761 gaagagaatc ctcgacaaga actacaagga caaggaacag aagaacttca aggatcggaa
     2821 cctgaacgac acccggtaca tcgccaggct ggtgttaaat tacaccaagg actacctgga
     2881 tttcctgccc ctgagcgacg acgagaacac caagctgaac gacacacaga agggcagcaa
     2941 ggtgcacgtg gaagccaaga gcggcatgct gaccagcgca ctgcgccaca cttggggctt
     3001 cagcgctaag gaccggaaca accacctgca tcacgccatc gatgccgtga tcatagccta
     3061 cgccaacaac tcaatcgtga aagctttcag tgactttaag aaagaacagg agagcaactc
     3121 tgccgaactg tacgccaaga aaattagcga gctggactac aagaacaaac ggaagttctt
     3181 cgaacctttc tcaggattta gacagaaggt gctggataag atcgatgaaa tcttcgtgtc
     3241 caagcccgag agaaagaagc ctagcggagc cctgcacaag gaaaccttca gaaaggaaga
     3301 agagttctac cagtcttatg ggggcaaaga aggcgtgctg aaggccctgg aactcggcaa
     3361 gatcagaaag gtgaagggaa agattgtgaa gaacggcgac atgttcagag tggacatctt
     3421 caagcacaag aagaccaaca agttctatgc cgtgcctatc tacacaatgg atttcgccct
     3481 gaaagtgctg cctaacaagg ctgtcgccag atctaagaag ggcgagatca aggactggat
     3541 cctgatggac gaaaactacg agttctgctt cagcctgtac aaggacagcc tgatcctgat
     3601 ccaaacaaag aagatgcagg agcctgaatt cgtgtactac aacgccttca caagcagcac
     3661 cgtgagcctg atcgtgtcta aacatgataa caagttcgaa accctgtcca agaaccaaaa
     3721 gatcctgttc aagaacgcca acgagaagga agtgatcgcc aagggcatcg gcattcagaa
     3781 tctgaaggtg ttcgagaaat atatcgtgtc cgctctggga gaggttacaa aggccgagtt
     3841 tcggcagaga gaagatttta agaagagcgg cgggagcggc gggagcgggg ggagcactaa
     3901 tctgagcgac atcattgaga aggagactgg gaaacagctg gtcattcagg agtccatcct
     3961 gatgctgcct gaggaggtgg aggaagtgat cggcaacaag ccagagtctg acatcctggt
     4021 gcacaccgcc tacgacgagt ccacagatga gaatgtgatg ctgctgacct ctgacgcccc
     4081 cgagtataag ccttgggccc tggtcatcca ggattctaac ggcgagaata agatcaagat
     4141 gctgagcgga ggatccggag gatctggagg cagcaccaac ctgtctgaca tcatcgagaa
     4201 ggagacaggc aagcagctgg tcatccagga gagcatcctg atgctgcccg aagaagtcga
     4261 agaagtgatc ggaaacaagc ctgagagcga tatcctggtc cataccgcct acgacgagag
     4321 taccgacgaa aatgtgatgc tgctgacatc cgacgcccca gagtataagc cctgggctct
     4381 ggtcatccag gattccaacg gagagaacaa aatcaaaatg ctgtctggcg gctcaaaaag
     4441 aaccgccgac ggcagcgaat tcgagcccaa gaagaagagg aaagtctaaa ccggtcatca
     4501 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     4561 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4621 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4681 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4741 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     4801 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4861 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     4921 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4981 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5041 tttgcgtatt gggcgctaca gacatgataa gatacattga tgagtttgga caaaccacaa
     5101 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5161 taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc
     5221 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     5281 tagaaggcgg cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc ctcggccacg
     5341 aagtgcttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt
     5401 cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac
     5461 ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat
     5521 acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga gttctgcaca
     5581 aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag
     5641 agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc
     5701 tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatggttt agttcctcac
     5761 cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga
     5821 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     5881 aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca
     5941 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     6001 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     6061 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     6121 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     6181 tgacacacat tccacagaat tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6241 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6301 aaagaacatg taagtgatct ccaaaaaata agtacttttt gactgtaaat aaaattgtaa
     6361 ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa gtaaaagtaa aagtattgat
     6421 ttttaattgt actcaagtaa agtaaaaatc cccaaaaata atacttaagt acagtaatca
     6481 agtaaaatta ctcaagtact ttacacctct gcatgtgagc aaaaggccag caaaaggcca
     6541 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6601 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6661 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6721 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6781 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6841 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6901 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6961 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7021 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7081 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7141 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacactcagt
     7201 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7261 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7321 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7381 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7441 catctggccc cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat
     7501 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7561 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7621 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7681 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7741 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7801 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7861 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7921 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7981 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8041 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8101 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8161 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8221 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8281 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8341 atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt
     8401 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     8461 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     8521 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgggtaccg gcatatggtt
     8581 cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac aagtacttag
     8641 ggaaaatttt actcaattaa aagtaaaagt atctggctag aatcttactt gagtaaaagt
     8701 aaaaaagtac tccattaaaa ttgtacttga gtattaacgc gttgacattg attattgact
     8761 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     8821 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     8881 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     8941 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     9001 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     9061 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     9121 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     9181 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     9241 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     9301 cggtgggagg tctatataag c
//
