LOCUS       sequence-470115-       13149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iSM_PPL2 prime editor and pegRNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13149)
  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 13149)
  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_PPL2
FEATURES             Location/Qualifiers
     source          1..13149
                     /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             8525..8545
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8555..8611
                     /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             8612..9208
                     /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(8612..8631)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9108..9128
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             9215..10210
                     /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(10232..10249)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10238..10462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   10646..10708
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      complement(10774..11362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10854..10873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11533..12324,12325..12393))
                     /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     12156..12175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12394..12498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12566..12584
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(12622..12644)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     12744..12763
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     repeat_region   12787..12821
                     /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 cccgtgctgg tggtggccaa
     5701 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaag cagctgatcc ccatcagcgt
     5761 gatgaacaag aaacagttcg agcagaaccc cgtgaagttc ctgcgggacc ggggctacca
     5821 gcaggtgggc aagaacgact tcatcaagct gcccaagtac accctggtgg acatcggcga
     5881 cggcatcaag cggctgtggg ccagcagcaa ggagatccac aagggcaacc agctggtggt
     5941 gagcaagaag agccagatcc tgctgtacca cgcccaccac ctggacagcg acctgagcaa
     6001 cgactacctg cagaaccaca accagcagtt cgacgtgctg ttcaacgaga tcatcagctt
     6061 cagcaaaaag tgcaagctgg gcaaggagca catccagaag atcgagaacg tgtacagcaa
     6121 caagaagaac agcgccagca tcgaggagct ggccgagagc tttatcaaac tgctcggctt
     6181 cacccagctg ggcgccacca gccccttcaa cttcctgggc gtgaagctga accagaagca
     6241 gtacaagggc aagaaggact acatcctgcc ctgcaccgag ggcaccctga tcagacagag
     6301 catcaccgga ctgtacgaaa ccagagtgga tctgagcaag atcggcgagg attctggagg
     6361 atctagcgga ggatcctctg gcagcgagac accaggaaca agcgagtcag caacaccaga
     6421 gagcagtggc ggcagcagcg gcggcagcag caccctaaat atagaagatg agtatcggct
     6481 acatgaaacc tcaaaagagc cagatgtttc tctagggtcc acatggctgt ctgattttcc
     6541 tcaggcctgg gcggaaaccg ggggcatggg actggcagtt cgccaagctc ctctgatcat
     6601 acctctgaaa gcaacctcta cccccgtgtc cataaaacaa taccccatgt cacaagaagc
     6661 cagactgggg atcaagcccc acatacagag actgttggac cagggaatac tggtaccctg
     6721 ccagtccccc tggaacacgc ccctgctacc cgttaagaaa ccagggacta atgattatag
     6781 gcctgtccag gatctgagag aagtcaacaa gcgggtggag gacatccacc ccaccgtgcc
     6841 caacccttac aacctcttga gcgggctccc accgtcccac cagtggtaca ctgtgcttga
     6901 tttaaaggat gcctttttct gcctgagact ccaccccacc agtcagcctc tcttcgcctt
     6961 tgagtggaga gatccagaga tgggaatctc aggacaattg acctggacca gactcccaca
     7021 gggtttcaaa aacagtccca ccctgtttaa tgaggcactg cacagggacc tagcagactt
     7081 cagaatccag cacccagact tgatcctgct acagtacgtg gatgacttac tgctggccgc
     7141 cacttctgag ctagactgcc aacaaggtac tcgggccctg ttacaaaccc tagggaacct
     7201 cgggtatcgg gcctcggcca agaaagccca aatttgccag aaacaggtca agtatctggg
     7261 gtatcttcta aaagagggtc agagatggct gactgaggcc agaaaagaga ctgtgatggg
     7321 gcagcctact cccaagaccc ctcgacaact aagggagttc ctagggaagg caggcttctg
     7381 tcgcctcttc atccctgggt ttgcagaaat ggcagccccc ctgtaccctc tcaccaaacc
     7441 ggggactctg tttaattggg gcccagacca acaaaaggcc tatcaagaaa tcaagcaagc
     7501 tcttctaact gccccagccc tggggttgcc agatttgact aagccctttg aactctttgt
     7561 cgacgagaag cagggctacg ccaaaggtgt cctaacgcaa aaactgggac cttggcgtcg
     7621 gccggtggcc tacctgtcca aaaagctaga cccagtagca gctgggtggc ccccttgcct
     7681 acggatggta gcagccattg ccgtactgac aaaggatgca ggcaagctaa ccatgggaca
     7741 gccactagtc attctggccc cccatgcagt agaggcacta gtcaaacaac cccccgaccg
     7801 ctggctttcc aacgcccgga tgactcacta tcaggccttg cttttggaca cggaccgggt
     7861 ccagttcggc ccggtggtag ccctgaaccc ggctacgctg ctcccactgc ctgaggaagg
     7921 gctgcaacac aactgccttg atatcctggc cgaagcccac ggaacccgac ccgacctaac
     7981 ggaccagccg ctcccagacg ccgaccacac ctggtacacg gatggaagca gtctcttaca
     8041 agagggacag cgtaaggcgg gagctgcggt gaccaccgag acagaggtaa tctgggctaa
     8101 agccctgcca gccgggacat ccgctcagcg ggctgaactg atagcactca cccaggccct
     8161 aaagatggca gaaggtaaga agctaaatgt ttatactgat agccgttatg cttttgctac
     8221 tgcccatatc catggagaaa tatacagaag gcgtgggtgg ctcacatcag aaggcaaaga
     8281 gatcaaaaat aaagacgaga tcttggccct actaaaagcc ctctttctgc ccaaaagact
     8341 tagcataatc cattgtccag gacatcaaaa gggacacagc gccgaggcta gaggcaaccg
     8401 gatggctgac caagcggccc gaaaggcagc catcacagag actccagaca cctctaccct
     8461 cctcatagaa aattcatcac cctctggcgg ctcaaaaaga accgccgacg gcagcgaatt
     8521 cgagcccaag aagaagagga aagtcggctc cggcgctacc aatttctccc tgctcaagca
     8581 agccggagac gtcgaggaaa acccaggccc aatgaccgag tacaagccca cggtgcgcct
     8641 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     8701 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     8761 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     8821 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     8881 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     8941 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     9001 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     9061 ggccgagcgc gccggggtgc ccgccttcct ggagacatcc gcgccccgca acctcccctt
     9121 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     9181 ctggtgcatg acccgcaagc ccggtgccgg ctccatgccc acgctactgc gggtttatat
     9241 agacggtcct cacgggatgg ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc
     9301 gcgcgacgat atcgtctacg tacccgagcc gatgacttac tggcaggtgc tgggggcttc
     9361 cgagacaatc gcgaacatct acaccacaca acaccgcctc gaccagggtg agatatcggc
     9421 cggggacgcg gcggtggtaa tgacaagcgc ccagataaca atgggcatgc cttatgccgt
     9481 gaccgacgcc gttctggctc ctcatatcgg gggggaggct gggagctcac atgccccgcc
     9541 cccggccctc accctcatct tcgaccgcca tcccatcgcc gccctcctgt gctacccggc
     9601 cgcgcgatac cttatgggca gcatgacccc ccaggccgtg ctggcgttcg tggccctcat
     9661 cccgccgacc ttgcccggca caaacatcgt gttgggggcc cttccggagg acagacacat
     9721 cgaccgcctg gccaaacgcc agcgccccgg cgagcggctt gacctggcta tgctggccgc
     9781 gattcgccgc gtttacgggc tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc
     9841 gtggcgggag gattggggac agctttcggg gacggccgtg ccgccccagg gtgccgagcc
     9901 ccagagcaac gcgggcccac gaccccatat cggggacacg ttatttaccc tgtttcgggc
     9961 ccccgagttg ctggccccca acggcgacct gtacaacgtg tttgcctggg ccttggacgt
    10021 cttggccaaa cgcctccgtc ccatgcacgt ctttatcctg gattacgacc aatcgcccgc
    10081 cggctgccgg gacgccctgc tgcaacttac ctccgggatg gtccagaccc acgtcaccac
    10141 ccccggctcc ataccgacga tctgcgacct ggcgcgcacg tttgcccggg agatggggga
    10201 ggctaactga gctctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
    10261 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
    10321 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
    10381 gggggtgggg tggggcagga cagcaagggg gaggattggg aggacaatag caggcatgct
    10441 ggggatgcgg tgggctctat ggggcgcgcc taaaagtttt gttactttat agaagaaatt
    10501 ttgagttttt gttttttttt aataaataaa taaacataaa taaattgttt gttgaattta
    10561 ttattagtat gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa
    10621 cctcgatata cagaccgata aaacacatgc gtcaatttta cgcatgatta tctttaacgt
    10681 acgtcacaat atgattatct ttctagggtt aaacatgtga gcaaaaggcc agcaaaaggc
    10741 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    10801 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    10861 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    10921 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    10981 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    11041 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    11101 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    11161 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    11221 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    11281 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    11341 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    11401 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    11461 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    11521 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    11581 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    11641 accatctggc cccagtgctg caatgatacc gcgactccca cgctcaccgg ctccagattt
    11701 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    11761 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    11821 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    11881 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    11941 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    12001 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    12061 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    12121 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    12181 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    12241 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    12301 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    12361 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    12421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    12481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
    12541 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
    12601 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
    12661 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
    12721 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccaatt
    12781 tgttaaccct agaaagatag tctgcgtaaa attgacgcat gcattcttga aatattgctc
    12841 tctctttcta aatagcgcga atccgtcgct gtgcatttag gacatctcag tcgccgcttg
    12901 gagctcccgt gaggcgtgct tgtcaatgcg gtaagtgtca ctgattttga actataacga
    12961 ccgcgtgagt caaaatgacg catgattatc ttttacgtga cttttaagat ttaactcata
    13021 cgataattat attgttattt catgttctac ttacgtgata acttattata tatatatttt
    13081 cttgttatag atatcatgta caaaaaagca ggctttaaag gaaccaattc agtcgactgg
    13141 acccggtac
//
