LOCUS       sequence-489519-       10471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9-SpG in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10471)
  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 10471)
  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: SpG-IP
FEATURES             Location/Qualifiers
     source          1..10471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1250..5353
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5408..5428
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5492..6044
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5636..5653)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5863..5882
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6056..6655
                     /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(6056..6075)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6552..6572
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6681..7269
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6734..6754)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7152..7163)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7341..7574
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7652..7773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7689..7708)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7743..7762
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7813..7948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7968..7987)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7969..7987)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7997..8014)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7997..8013)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8120..8137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8291..8879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8371..8390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9050..9841,9842..9910))
                     /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     9673..9692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9911..10015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10094..10113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    10247..10318
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(10323..10369)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    10387..10435
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 ctagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcgt ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgtggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat cagcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg cccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagcac aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcca gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa aactacccct aaaagccaaa
     1201 agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgca tggacaagaa
     1261 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     1321 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     1381 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     1441 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     1501 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     1561 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     1621 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     1681 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     1741 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     1801 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     1861 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     1921 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     1981 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     2041 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     2101 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     2161 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     2221 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     2281 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     2341 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     2401 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     2461 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     2521 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     2581 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     2641 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     2701 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     2761 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     2821 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     2881 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     2941 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     3001 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     3061 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     3121 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     3181 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     3241 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     3301 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     3361 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     3421 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     3481 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     3541 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     3601 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     3661 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     3721 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
     3781 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
     3841 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     3901 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
     3961 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     4021 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     4081 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     4141 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     4201 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     4261 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     4321 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     4381 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     4441 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     4501 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     4561 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     4621 ggactgggac cctaagaagt acggcggctt cctgtggccc accgtggcct attctgtgct
     4681 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     4741 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     4801 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     4861 cgagctggaa aacggccgga agagaatgct ggcctctgcc aagcagctgc agaagggaaa
     4921 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     4981 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     5041 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     5101 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     5161 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     5221 caagtacttt gacaccacca tcgaccggaa gcagtacaga agcaccaaag aggtgctgga
     5281 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     5341 gctgggaggt gactctggag gatctggagg cagcaagaga accgccgatg gaagcgagtt
     5401 cgagtctccc aagaagaaga ggaaagtcgg aagcggataa gctgccaagc ttccgagctc
     5461 tcgaattcaa aggaggtacc ccccccccct aacgttactg gccgaagccg cttggaataa
     5521 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     5581 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     5641 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     5701 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     5761 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     5821 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     5881 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     5941 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     6001 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatgac
     6061 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     6121 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     6181 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     6241 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     6301 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     6361 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     6421 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     6481 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     6541 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     6601 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaaaacc
     6661 cgctgatcag caaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     6721 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     6781 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     6841 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6901 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     6961 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     7021 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     7081 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     7141 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     7201 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     7261 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     7321 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     7381 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     7441 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     7501 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     7561 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
     7621 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     7681 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     7741 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     7801 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     7861 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     7921 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     7981 cgtattacgc gcgctcactg gccgtcgttt tacactcact gcccgctttc cagtcgggaa
     8041 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc gggaagaggc ggtttgcgta
     8101 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     8161 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     8221 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     8281 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     8341 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8401 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8461 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8521 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     8581 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8641 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8701 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8761 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8821 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8881 aagatccttt gatcttttct acggggtctg acactcagtg gaacgaaaac tcacgttaag
     8941 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     9001 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     9061 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     9121 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     9181 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     9241 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     9301 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     9361 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     9421 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     9481 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     9541 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9601 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9661 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9721 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9781 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9841 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9901 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9961 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    10021 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag
    10081 ggtcgactct cagtacaatc tgctctgatg ccgctcgact ctggctctat cgaatctccg
    10141 tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg
    10201 tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt
    10261 agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa
    10321 ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat caatcgttgc
    10381 gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt ttattatcta
    10441 actgctgatc gagtgtagcc agatctcgtg a
//
