LOCUS       sequence-343635-       12075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of SpCas9 prime editor 2 with P2A-human MLH1dn 
            (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12075)
  AUTHORS   Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen
            C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, 
            Liu DR
  TITLE     Enhanced prime editing systems by manipulating cellular determinants
            of editing outcomes.
  JOURNAL   Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 
            10.1016/j.cell.2021.09.018.
  PUBMED    34653350
REFERENCE   2  (bases 1 to 12075)
  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"; doi: 
            "10.1016/j.cell.2021.09"; journalName: "Cell"; date: "2021-10-28- 
            28"; volume: "184"; issue: "22"; pages: "5635-5652.e29"
COMMENT     Sequence Label: pCMV-PE2-P2A-hMLH1dn
FEATURES             Location/Qualifiers
     source          1..12075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        519..722
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     673..693
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     764..783
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        764..782
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             832..852
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(853..3366,3367..3369,3370..4953)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             853..4953
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7126..7146
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7156..7212
                     /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             9477..9494
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9517..9534)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9523..9747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9818..9834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9818..9834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9831..9853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9842..9858
                     /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(9866..9872,9873..9890,9891..9896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9911..9932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10049..10066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10220..10808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10300..10319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10979..11770,11771..11839))
                     /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     11602..11621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11840..11944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12023..12042)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
       61 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      121 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      181 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      241 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      301 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      361 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      421 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      481 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      541 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      601 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      661 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      721 ctggtttagt gaaccgtcag atccgctaga gatccgcggc cgctaatacg actcactata
      781 gggagagccg ccaccatgaa acggacagcc gacggaagcg agttcgagtc accaaagaag
      841 aagcggaaag tcgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
      901 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
      961 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     1021 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     1081 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     1141 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     1201 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     1261 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     1321 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     1381 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     1441 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     1501 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     1561 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     1621 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     1681 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     1741 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     1801 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     1861 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     1921 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     1981 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     2041 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     2101 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     2161 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     2221 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     2281 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     2341 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     2401 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     2461 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     2521 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     2581 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     2641 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     2701 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     2761 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     2821 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     2881 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     2941 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     3001 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     3061 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     3121 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     3181 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     3241 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     3301 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     3361 gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     3421 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     3481 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     3541 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     3601 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     3661 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     3721 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     3781 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     3841 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     3901 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     3961 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     4021 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     4081 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     4141 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     4201 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     4261 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     4321 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     4381 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     4441 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     4501 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     4561 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     4621 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     4681 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     4741 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     4801 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     4861 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     4921 acacggatcg acctgtctca gctgggaggt gactctggag gatctagcgg aggatcctct
     4981 ggcagcgaga caccaggaac aagcgagtca gcaacaccag agagcagtgg cggcagcagc
     5041 ggcggcagca gcaccctaaa tatagaagat gagtatcggc tacatgagac ctcaaaagag
     5101 ccagatgttt ctctagggtc cacatggctg tctgattttc ctcaggcctg ggcggaaacc
     5161 gggggcatgg gactggcagt tcgccaagct cctctgatca tacctctgaa agcaacctct
     5221 acccccgtgt ccataaaaca ataccccatg tcacaagaag ccagactggg gatcaagccc
     5281 cacatacaga gactgttgga ccagggaata ctggtaccct gccagtcccc ctggaacacg
     5341 cccctgctac ccgttaagaa accagggact aatgattata ggcctgtcca ggatctgaga
     5401 gaagtcaaca agcgggtgga agacatccac cccaccgtgc ccaaccctta caacctcttg
     5461 agcgggctcc caccgtccca ccagtggtac actgtgcttg atttaaagga tgcctttttc
     5521 tgcctgagac tccaccccac cagtcagcct ctcttcgcct ttgagtggag agatccagag
     5581 atgggaatct caggacaatt gacctggacc agactcccac agggtttcaa aaacagtccc
     5641 accctgttta atgaggcact gcacagagac ctagcagact tccggatcca gcacccagac
     5701 ttgatcctgc tacagtacgt ggatgactta ctgctggccg ccacttctga gctagactgc
     5761 caacaaggta ctcgggccct gttacaaacc ctagggaacc tcgggtatcg ggcctcggcc
     5821 aagaaagccc aaatttgcca gaaacaggtc aagtatctgg ggtatcttct aaaagagggt
     5881 cagagatggc tgactgaggc cagaaaagag actgtgatgg ggcagcctac tccgaagacc
     5941 cctcgacaac taagggagtt cctagggaag gcaggcttct gtcgcctctt catccctggg
     6001 tttgcagaaa tggcagcccc cctgtaccct ctcaccaaac cggggactct gtttaattgg
     6061 ggcccagacc aacaaaaggc ctatcaagaa atcaagcaag ctcttctaac tgccccagcc
     6121 ctggggttgc cagatttgac taagcccttt gaactctttg tcgacgagaa gcagggctac
     6181 gccaaaggtg tcctaacgca aaaactggga ccttggcgtc ggccggtggc ctacctgtcc
     6241 aaaaagctag acccagtagc agctgggtgg cccccttgcc tacggatggt agcagccatt
     6301 gccgtactga caaaggatgc aggcaagcta accatgggac agccactagt cattctggcc
     6361 ccccatgcag tagaggcact agtcaaacaa ccccccgacc gctggctttc caacgcccgg
     6421 atgactcact atcaggcctt gcttttggac acggaccggg tccagttcgg accggtggta
     6481 gccctgaacc cggctacgct gctcccactg cctgaggaag ggctgcaaca caactgcctt
     6541 gatatcctgg ccgaagccca cggaacccga cccgacctaa cggaccagcc gctcccagac
     6601 gccgaccaca cctggtacac ggatggaagc agtctcttac aagagggaca gcgtaaggcg
     6661 ggagctgcgg tgaccaccga gaccgaggta atctgggcta aagccctgcc agccgggaca
     6721 tccgctcagc gggctgaact gatagcactc acccaggccc taaagatggc agaaggtaag
     6781 aagctaaatg tttatactga tagccgttat gcttttgcta ctgcccatat ccatggagaa
     6841 atatacagaa ggcgtgggtg gctcacatca gaaggcaaag agatcaaaaa taaagacgag
     6901 atcttggccc tactaaaagc cctctttctg cccaaaagac ttagcataat ccattgtcca
     6961 ggacatcaaa agggacacag cgccgaggct agaggcaacc ggatggctga ccaagcggcc
     7021 cgaaaggcag ccatcacaga gactccagac acctctaccc tcctcataga aaattcatca
     7081 ccctctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa gaagaagagg
     7141 aaagtcggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
     7201 aaccctggac ctagcttcgt tgctggagtc atccggagac tggacgagac agtggtgaac
     7261 agaattgccg ccggcgaggt gatccagaga cctgccaatg caataaagga gatgatcgag
     7321 aactgtctgg acgccaagtc cacaagcatt caggtgatcg tgaaggaggg cggactgaag
     7381 ctgatccaga tccaagacaa cggcacaggc atcagaaagg aagatctgga catcgtgtgt
     7441 gaacggttca ccacatctaa gctgcagtct tttgaggatc tggcctctat cagtacctac
     7501 ggcttcagag gcgaggccct ggccagcatc agccacgtgg cccatgtgac catcaccacc
     7561 aaaaccgccg acggcaaatg cgcttatcgc gctagctaca gcgacggcaa gctgaaagcc
     7621 ccgccaaagc cttgcgccgg caaccagggt acacagataa cagtggagga tctgttctac
     7681 aacatcgcca cccggagaaa ggccctgaaa aatcccagcg aggagtacgg caagatcctg
     7741 gaagtcgtcg gcagatactc cgtgcacaac gccggaatca gctttagcgt aaagaagcag
     7801 ggagaaaccg tggccgatgt gcgcaccctg ccaaatgcca gcaccgtgga taacatcaga
     7861 agcattttcg gaaatgccgt gtccagagaa ctgatcgaga tcggctgcga agataagacc
     7921 ctggctttta agatgaacgg ctacatcagc aacgccaatt actctgtgaa gaagtgcatc
     7981 tttcttctgt tcatcaacca cagactggtg gaaagcacca gcctgcggaa agccatcgag
     8041 acagtgtacg ccgcctacct gcctaagaac acccacccct tcctgtacct gagcctcgag
     8101 atcagccctc agaacgtgga cgtcaatgtg catcctacaa agcacgaggt gcacttcctg
     8161 cacgaggaaa gcatcctgga aagagtgcag cagcacattg agagcaagct gctgggctct
     8221 aacagcagca gaatgtactt cacacagacc ctgctgcctg gcctggccgg cccctcaggc
     8281 gaaatggtta agtccacaac ctctctgacc tcatccagca ccagcggttc ttccgataag
     8341 gtgtacgccc accagatggt gcggaccgac tctcgggagc agaagctgga cgcctttctg
     8401 caacctctga gcaaacctct gagctctcag cctcaggcca tcgtgaccga ggacaagaca
     8461 gatatctcct ccggccgtgc cagacagcag gacgaagaaa tgctcgagct gccagctcct
     8521 gccgaggtgg ccgccaagaa ccagagcctg gagggagata ccacaaaggg caccagcgaa
     8581 atgagcgaga agcggggccc tacctccagc aaccccagaa aacggcaccg ggaggacagc
     8641 gacgtggaaa tggtggagga cgacagcaga aaggaaatga cagccgcttg tacccctaga
     8701 agaagaatca tcaacctgac ctccgtgctg agcctgcagg aggagatcaa cgagcagggc
     8761 cacgaggtgc tgagagagat gctgcacaat cacagcttcg tgggctgcgt gaaccctcaa
     8821 tgggccctgg ctcagcatca aacaaagctg tacctgctga acaccaccaa gctgagcgaa
     8881 gagctgttct accagatcct catctacgac ttcgccaact tcggcgtgct acgcctgagc
     8941 gagcccgccc ctctgtttga cctggccatg ctggctctgg atagcccaga aagcggctgg
     9001 acagaagagg acggacctaa agaggggctg gctgaataca tcgtggagtt cctgaagaaa
     9061 aaggccgaga tgctggccga ctacttttct ctggaaatcg acgaggaagg caacctgatc
     9121 ggcctgcctc tgctgatcga taactacgtg cctcccctgg aaggcctgcc catcttcatc
     9181 ctgagactgg ctacagaggt gaactgggac gaggaaaagg aatgcttcga gtctctgagc
     9241 aaggagtgcg ccatgttcta tagcatcaga aaacagtaca tctctgaaga gagcactctg
     9301 tctggccagc agagtgaagt gcccggaagc atccccaaca gctggaagtg gaccgtggaa
     9361 cacatcgtgt acaaggccct gcggagccac attctccctc ctaagcactt caccgaggac
     9421 ggcaacatcc tgcagctggc caacctgccc gacctttata aggttttcta accggtcatc
     9481 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     9541 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     9601 ctgtcctttc ctaataaaat gagaaaattg catcgcattg tctgagtagg tgtcattcta
     9661 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     9721 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
     9781 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     9841 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     9901 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     9961 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    10021 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    10081 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    10141 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    10201 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    10261 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    10321 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    10381 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    10441 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    10501 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    10561 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    10621 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
    10681 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    10741 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    10801 gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg aacgaaaact
    10861 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
    10921 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
    10981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
    11041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
    11101 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
    11161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
    11221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    11281 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    11341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    11401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    11461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    11521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    11581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    11641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    11701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    11761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    11821 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    11881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    11941 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
    12001 atcccctagg gtctactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
    12061 tgctccctgc ttgtg
//
