LOCUS       sequence-470114-       13116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iSM (iSpyMac) prime editor and pegRNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13116)
  AUTHORS   Eggenschwiler R, Hoffmann T, Dmytrenko O, Opitz M, Ackel-Zakour M, 
            Wang P, McCallan SA, Fraguas-Eggenschwiler M, Niemann H, Patronov A,
            Beisel CL, Cantz T
  TITLE     PAM-interacting domain turn-helix 51 motifs can improve Cas9-SpRY 
            activity.
  JOURNAL   Nucleic Acids Res. 2025 Aug 11;53(15):gkaf782. doi: 
            10.1093/nar/gkaf782.
  PUBMED    40808297
REFERENCE   2  (bases 1 to 13116)
  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: "Nucleic 
            Acids Res."; date: "2025-08-11"; volume: "53(15):gkaf782. doi"; 
            pages: " 10.1093/nar/gkaf782"
COMMENT     Sequence Label: PB-PE iSM
FEATURES             Location/Qualifiers
     source          1..13116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..265
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     17..37
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     188..207
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        294..369
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        415..794
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        796..1071
                     /label=chicken beta-actin promoter
     intron          1072..2088
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2012..2034
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2096..2115
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2154..2173
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2154..2172
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2222..2242
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2243..3958
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             8492..8512
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8522..8578
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8579..9175
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8579..8598)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9075..9095
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             9182..10177
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(10199..10216)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10205..10429
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   10613..10675
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      complement(10741..11329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10821..10840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11500..12291,12292..12360))
                     /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     12123..12142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12361..12465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12533..12551
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(12589..12611)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     12711..12730
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     repeat_region   12754..12788
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 caaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata
       61 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
      121 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      181 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      241 tatcttgtgg aaaggacgaa acaccgggtc ttcgtggagc tcatcgaaga cctgttttag
      301 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      361 agtcggtgca gagaccgaat cgaccagagg tctcgttttt ttctagacgc tagcgacatt
      421 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      481 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      541 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      601 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      661 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      721 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      781 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
      841 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
      901 cggggggggg gggggggcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
      961 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     1021 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     1081 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     1141 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     1201 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
     1261 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     1321 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     1381 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     1441 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
     1501 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
     1561 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     1621 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     1681 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     1741 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     1801 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     1861 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1921 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     1981 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     2041 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     2101 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgatagcggc cgctaatacg
     2161 actcactata gggagagccg ccaccatgaa acggacagcc gacggaagcg agttcgagtc
     2221 accaaagaag aagcggaaag tcgacaagaa gtacagcatc ggcctggaca tcggcaccaa
     2281 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     2341 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     2401 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     2461 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     2521 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     2581 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     2641 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     2701 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     2761 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     2821 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     2881 cagactgagc aagagcagaa agctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     2941 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     3001 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     3061 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     3121 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     3181 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     3241 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     3301 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     3361 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     3421 gagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     3481 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     3541 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     3601 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     3661 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     3721 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     3781 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     3841 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     3901 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     3961 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     4021 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     4081 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     4141 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     4201 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     4261 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     4321 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     4381 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     4441 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     4501 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     4561 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     4621 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     4681 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     4741 cgactacgat gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     4801 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     4861 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     4921 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     4981 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     5041 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     5101 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     5161 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctaaacg ccgtcgtggg
     5221 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     5281 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     5341 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     5401 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     5461 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     5521 cgtgaaaaag accgagatcc agaccgtggg acagaatgga ggcctgttcg acgacaaccc
     5581 caagagcccc ctggaggtga cccccagcaa actggtgccc ctgaagaaag agctgaaccc
     5641 caagaaatac ggcggctacc agaagcccac caccgcctac cccgtgctgc tgatcaccga
     5701 caccaagcag ctgatcccca tcagcgtgat gaacaagaaa cagttcgagc agaaccccgt
     5761 gaagttcctg cgggaccggg gctaccagca ggtgggcaag aacgacttca tcaagctgcc
     5821 caagtacacc ctggtggaca tcggcgacgg catcaagcgg ctgtgggcca gcagcaagga
     5881 gatccacaag ggcaaccagc tggtggtgag caagaagagc cagatcctgc tgtaccacgc
     5941 ccaccacctg gacagcgacc tgagcaacga ctacctgcag aaccacaacc agcagttcga
     6001 cgtgctgttc aacgagatca tcagcttcag caaaaagtgc aagctgggca aggagcacat
     6061 ccagaagatc gagaacgtgt acagcaacaa gaagaacagc gccagcatcg aggagctggc
     6121 cgagagcttt atcaaactgc tcggcttcac ccagctgggc gccaccagcc ccttcaactt
     6181 cctgggcgtg aagctgaacc agaagcagta caagggcaag aaggactaca tcctgccctg
     6241 caccgagggc accctgatca gacagagcat caccggactg tacgaaacca gagtggatct
     6301 gagcaagatc ggcgaggatt ctggaggatc tagcggagga tcctctggca gcgagacacc
     6361 aggaacaagc gagtcagcaa caccagagag cagtggcggc agcagcggcg gcagcagcac
     6421 cctaaatata gaagatgagt atcggctaca tgaaacctca aaagagccag atgtttctct
     6481 agggtccaca tggctgtctg attttcctca ggcctgggcg gaaaccgggg gcatgggact
     6541 ggcagttcgc caagctcctc tgatcatacc tctgaaagca acctctaccc ccgtgtccat
     6601 aaaacaatac cccatgtcac aagaagccag actggggatc aagccccaca tacagagact
     6661 gttggaccag ggaatactgg taccctgcca gtccccctgg aacacgcccc tgctacccgt
     6721 taagaaacca gggactaatg attataggcc tgtccaggat ctgagagaag tcaacaagcg
     6781 ggtggaggac atccacccca ccgtgcccaa cccttacaac ctcttgagcg ggctcccacc
     6841 gtcccaccag tggtacactg tgcttgattt aaaggatgcc tttttctgcc tgagactcca
     6901 ccccaccagt cagcctctct tcgcctttga gtggagagat ccagagatgg gaatctcagg
     6961 acaattgacc tggaccagac tcccacaggg tttcaaaaac agtcccaccc tgtttaatga
     7021 ggcactgcac agggacctag cagacttcag aatccagcac ccagacttga tcctgctaca
     7081 gtacgtggat gacttactgc tggccgccac ttctgagcta gactgccaac aaggtactcg
     7141 ggccctgtta caaaccctag ggaacctcgg gtatcgggcc tcggccaaga aagcccaaat
     7201 ttgccagaaa caggtcaagt atctggggta tcttctaaaa gagggtcaga gatggctgac
     7261 tgaggccaga aaagagactg tgatggggca gcctactccc aagacccctc gacaactaag
     7321 ggagttccta gggaaggcag gcttctgtcg cctcttcatc cctgggtttg cagaaatggc
     7381 agcccccctg taccctctca ccaaaccggg gactctgttt aattggggcc cagaccaaca
     7441 aaaggcctat caagaaatca agcaagctct tctaactgcc ccagccctgg ggttgccaga
     7501 tttgactaag ccctttgaac tctttgtcga cgagaagcag ggctacgcca aaggtgtcct
     7561 aacgcaaaaa ctgggacctt ggcgtcggcc ggtggcctac ctgtccaaaa agctagaccc
     7621 agtagcagct gggtggcccc cttgcctacg gatggtagca gccattgccg tactgacaaa
     7681 ggatgcaggc aagctaacca tgggacagcc actagtcatt ctggcccccc atgcagtaga
     7741 ggcactagtc aaacaacccc ccgaccgctg gctttccaac gcccggatga ctcactatca
     7801 ggccttgctt ttggacacgg accgggtcca gttcggcccg gtggtagccc tgaacccggc
     7861 tacgctgctc ccactgcctg aggaagggct gcaacacaac tgccttgata tcctggccga
     7921 agcccacgga acccgacccg acctaacgga ccagccgctc ccagacgccg accacacctg
     7981 gtacacggat ggaagcagtc tcttacaaga gggacagcgt aaggcgggag ctgcggtgac
     8041 caccgagaca gaggtaatct gggctaaagc cctgccagcc gggacatccg ctcagcgggc
     8101 tgaactgata gcactcaccc aggccctaaa gatggcagaa ggtaagaagc taaatgttta
     8161 tactgatagc cgttatgctt ttgctactgc ccatatccat ggagaaatat acagaaggcg
     8221 tgggtggctc acatcagaag gcaaagagat caaaaataaa gacgagatct tggccctact
     8281 aaaagccctc tttctgccca aaagacttag cataatccat tgtccaggac atcaaaaggg
     8341 acacagcgcc gaggctagag gcaaccggat ggctgaccaa gcggcccgaa aggcagccat
     8401 cacagagact ccagacacct ctaccctcct catagaaaat tcatcaccct ctggcggctc
     8461 aaaaagaacc gccgacggca gcgaattcga gcccaagaag aagaggaaag tcggctccgg
     8521 cgctaccaat ttctccctgc tcaagcaagc cggagacgtc gaggaaaacc caggcccaat
     8581 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     8641 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     8701 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     8761 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     8821 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     8881 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     8941 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     9001 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     9061 gacatccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     9121 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgccggctc
     9181 catgcccacg ctactgcggg tttatataga cggtcctcac gggatgggga aaaccaccac
     9241 cacgcaactg ctggtggccc tgggttcgcg cgacgatatc gtctacgtac ccgagccgat
     9301 gacttactgg caggtgctgg gggcttccga gacaatcgcg aacatctaca ccacacaaca
     9361 ccgcctcgac cagggtgaga tatcggccgg ggacgcggcg gtggtaatga caagcgccca
     9421 gataacaatg ggcatgcctt atgccgtgac cgacgccgtt ctggctcctc atatcggggg
     9481 ggaggctggg agctcacatg ccccgccccc ggccctcacc ctcatcttcg accgccatcc
     9541 catcgccgcc ctcctgtgct acccggccgc gcgatacctt atgggcagca tgacccccca
     9601 ggccgtgctg gcgttcgtgg ccctcatccc gccgaccttg cccggcacaa acatcgtgtt
     9661 gggggccctt ccggaggaca gacacatcga ccgcctggcc aaacgccagc gccccggcga
     9721 gcggcttgac ctggctatgc tggccgcgat tcgccgcgtt tacgggctgc ttgccaatac
     9781 ggtgcggtat ctgcagggcg gcgggtcgtg gcgggaggat tggggacagc tttcggggac
     9841 ggccgtgccg ccccagggtg ccgagcccca gagcaacgcg ggcccacgac cccatatcgg
     9901 ggacacgtta tttaccctgt ttcgggcccc cgagttgctg gcccccaacg gcgacctgta
     9961 caacgtgttt gcctgggcct tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt
    10021 tatcctggat tacgaccaat cgcccgccgg ctgccgggac gccctgctgc aacttacctc
    10081 cgggatggtc cagacccacg tcaccacccc cggctccata ccgacgatct gcgacctggc
    10141 gcgcacgttt gcccgggaga tgggggaggc taactgagct ctagagctcg ctgatcagcc
    10201 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
    10261 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
    10321 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
    10381 gattgggagg acaatagcag gcatgctggg gatgcggtgg gctctatggg gcgcgcctaa
    10441 aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat aaataaataa
    10501 acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact
    10561 taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc
    10621 aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaaa
    10681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    10741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    10801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    10861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    10921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    10981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    11041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    11101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    11161 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    11221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    11281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    11341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    11401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    11461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    11521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    11581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    11641 actcccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    11701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    11761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    11821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    11881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    11941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    12001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    12061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    12121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    12181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    12241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    12301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    12361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    12421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    12481 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
    12541 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
    12601 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
    12661 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
    12721 gcagattgta ctgagagtgc accaatttgt taaccctaga aagatagtct gcgtaaaatt
    12781 gacgcatgca ttcttgaaat attgctctct ctttctaaat agcgcgaatc cgtcgctgtg
    12841 catttaggac atctcagtcg ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta
    12901 agtgtcactg attttgaact ataacgaccg cgtgagtcaa aatgacgcat gattatcttt
    12961 tacgtgactt ttaagattta actcatacga taattatatt gttatttcat gttctactta
    13021 cgtgataact tattatatat atattttctt gttatagata tcatgtacaa aaaagcaggc
    13081 tttaaaggaa ccaattcagt cgactggacc cggtac
//
