LOCUS       sequence-338127-       11741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Delivers all prime editing nickase components in a single, puromycin
            selectable plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11741)
  AUTHORS   Adikusuma F, Lushington C, Arudkumar J, Godahewa GI, Chey YCJ, 
            Gierus L, Piltz S, Geiger A, Jain Y, Reti D, Wilson LOW, Bauer DC, 
            Thomas PQ
  TITLE     Optimized nickase- and nuclease-based prime editing in human and 
            mouse cells.
  JOURNAL   Nucleic Acids Res. 2021 Sep 17. pii: 6371974. doi: 
            10.1093/nar/gkab792.
  PUBMED    34534334
REFERENCE   2  (bases 1 to 11741)
  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. 2021 Sep 17. pii: 6371974. doi: 10.1093/nar/gkab792."
COMMENT     Sequence Label: PEA1-Puro
FEATURES             Location/Qualifiers
     source          1..11741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2886..2933
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2949..3002
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3003..3602
                     /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(3003..3022)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3499..3519
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3630..3647)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3636..3843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3852..3868
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3852..3868
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        complement(3911..3986)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4004..4252)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     repeat_region   4261..4401
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4261..4390
                     /label=AAV2 ITR
     rep_origin      4476..4931
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4563..4582)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4773..4794
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4948..4967)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5067..5089
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5127..5145)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5213..5317
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5318..5386,5387..6178)
                     /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     complement(5536..5555)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6349..6937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6838..6857
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6999..7247
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7266..7341
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        7426..7711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          7989..8216
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      8228..8237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8237..8302
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8309..8329
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(8354..10867,10868..10870,10871..11741,1..713)
                     /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             join(8354..11741,1..713)
                     /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"
ORIGIN
        1 acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa
       61 agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa
      121 gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga
      181 aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga
      241 agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca
      301 aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg
      361 ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg
      421 agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc
      481 tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc
      541 acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca
      601 tcgaccggaa gaggtacacc agcacgaaag aggtgctgga cgccaccctg atccaccaga
      661 gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggt gactctggag
      721 gatctagcgg aggatcctct ggcagcgaga caccaggaac aagcgagtca gcaacaccag
      781 agagcagtgg cggcagcagc ggcggcagca gcaccctaaa tatagaagat gagtatcggc
      841 tacatgagac ctcaaaagag ccagatgttt ctctagggtc cacatggctg tctgattttc
      901 ctcaggcctg ggcggaaacc gggggcatgg gactggcagt tcgccaagct cctctgatca
      961 tacctctgaa agcaacctct acccccgtgt ccataaaaca ataccccatg tcacaagaag
     1021 ccagactggg gatcaagccc cacatacaga gactgttgga ccagggaata ctggtaccct
     1081 gccagtcccc ctggaacacg cccctgctac ccgttaagaa accagggact aatgattata
     1141 ggcctgtcca ggatctgaga gaagtcaaca agcgggtgga ggacatccac cccaccgtgc
     1201 ccaaccctta caacctcttg agcgggctcc caccgtccca ccagtggtac actgtgcttg
     1261 atttaaagga tgcctttttc tgcctgagac tccaccccac cagtcagcct ctcttcgcct
     1321 ttgagtggag agatccagag atgggaatct caggacaatt gacctggacc agactcccac
     1381 agggtttcaa aaacagtccc accctgttta atgaggcact gcacagagac ctagcagact
     1441 tccggatcca gcacccagac ttgatcctgc tacagtacgt ggatgactta ctgctggccg
     1501 ccacttctga gctagactgc caacaaggta ctcgggccct gttacaaacc ctagggaacc
     1561 tcgggtatcg ggcctcggcc aagaaagccc aaatttgcca gaaacaggtc aagtatctgg
     1621 ggtatcttct aaaagagggt cagagatggc tgactgaggc cagaaaagag actgtgatgg
     1681 ggcagcctac tccaaagacc cctcgacaac taagggagtt cctagggaag gcaggcttct
     1741 gtcgcctctt catccctggg tttgcagaaa tggcagcccc cctgtaccct ctcaccaaac
     1801 cggggactct gtttaattgg ggcccagacc aacaaaaggc ctatcaagaa atcaagcaag
     1861 cccttctaac tgccccagcc ctggggttgc cagatttgac taagcccttt gaactctttg
     1921 tcgacgagaa gcagggctac gccaaaggtg tcctaacgca aaaactggga ccttggcgtc
     1981 ggccggtggc ctacctgtcc aaaaagctag acccagtagc agctgggtgg cccccttgcc
     2041 tacggatggt agcagccatt gccgtactga caaaggatgc aggcaagcta accatgggac
     2101 agccactagt cattctggcc ccccatgcag tagaggcact agtcaaacaa ccccccgacc
     2161 gctggctttc caacgcccgg atgactcact atcaggcctt gcttttggac acggaccggg
     2221 tccagttcgg accggtggta gccctgaacc cggctacgct gctcccactg cctgaggaag
     2281 ggctgcaaca caactgcctt gatatcctgg ccgaagccca cggaacccga cccgacctaa
     2341 cggaccagcc gctcccagac gccgaccaca cctggtacac ggatggaagc agtctcttac
     2401 aagagggaca gcgtaaggcg ggagctgcgg tgaccaccga gaccgaggta atctgggcta
     2461 aagccctgcc agccgggaca tccgctcagc gggctgaact gatagcactc acccaggccc
     2521 taaagatggc agaaggtaag aagctaaatg tttatactga tagccgttat gcttttgcta
     2581 ctgcccatat ccatggagaa atatacagaa ggcgtgggtg gctcacatca gaaggcaaag
     2641 agatcaaaaa taaagacgag atcttggccc tactaaaagc cctctttctg cccaaaagac
     2701 ttagcataat ccattgtcca ggacatcaaa agggacacag cgccgaggct agaggcaacc
     2761 ggatggctga ccaagcggcc cgaaaggcag ccatcacaga gactccagac acctctaccc
     2821 tcctcataga aaattcatca ccctctggcg gctcaaaaag aaccgccgac ggcagcgaat
     2881 tcgagaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggaattcg
     2941 gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
     3001 caatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     3061 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     3121 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     3181 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     3241 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     3301 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     3361 agcccgcgtg gttcctggcc accgtcggag tctcgcccga ccaccagggc aagggtctgg
     3421 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     3481 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     3541 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     3601 gagaattcta actagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3661 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3721 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3781 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg
     3841 ggagcggccg ctctagaact agtggatccc ccgggctgca ggaattcgat acgaatcctg
     3901 caggaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt
     3961 taacttgcta tttctagctc taaaacgtct tctcgaagac cccggtgttt cgtcctttcc
     4021 acaagatata taaagccaag aaatcgaaat actttcaagt tacggtaagc atatgatagt
     4081 ccattttaaa acataatttt aaaactgcaa actacccaag aaattattac tttctacgtc
     4141 acgtattttg tactaatatc tttgtgttta cagtcaaatt aattccaatt atctctctaa
     4201 cagccttgta tcgtatatgc aaatatgaag gaatcatggg aaataggccc tcgcggccgc
     4261 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4321 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4381 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4441 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4501 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     4561 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4621 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4681 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4741 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4801 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4861 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4921 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4981 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5041 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5101 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5161 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5221 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5281 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5341 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5401 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5461 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5521 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5581 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5641 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5701 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5761 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5821 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5881 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5941 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6001 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
     6061 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6121 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6181 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6241 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6301 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6361 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6421 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6481 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6541 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6601 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6661 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6721 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6781 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6841 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6901 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6961 acggttcctg gccttttgct ggccttttgc tcacatgtga gggcctattt cccatgattc
     7021 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
     7081 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     7141 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     7201 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggg tcttcgagaa
     7261 gacctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     7321 aaagtggcac cgagtcggtg cgggtcttcg agaagacctt ttttttgttc tagtccgttt
     7381 ttagcgcgtg cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac
     7441 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac
     7501 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     7561 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     7621 atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta
     7681 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
     7741 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
     7801 ttgtgcagcg atgggggcgg gggggggggg ggnnsgsgcc rggsggggsg gggsggggsg
     7861 rggggsgggg sggggsgrgg sggrgrggkg sggsggcagc caatcagagc ggcgcgctcc
     7921 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
     7981 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
     8041 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
     8101 cttctcctcc gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg
     8161 gtggggtatt aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg
     8221 gaccggtgcc accatggact ataaggacca cgacggagac tacaaggatc atgatattga
     8281 ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg
     8341 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     8401 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     8461 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     8521 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     8581 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     8641 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     8701 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     8761 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     8821 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     8881 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     8941 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     9001 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     9061 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     9121 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     9181 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     9241 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     9301 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     9361 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     9421 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     9481 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     9541 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     9601 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     9661 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     9721 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     9781 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     9841 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     9901 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     9961 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
    10021 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
    10081 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
    10141 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
    10201 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
    10261 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
    10321 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
    10381 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
    10441 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
    10501 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
    10561 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
    10621 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
    10681 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
    10741 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
    10801 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
    10861 tgtggacgct atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
    10921 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
    10981 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
    11041 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
    11101 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
    11161 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
    11221 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
    11281 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
    11341 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
    11401 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
    11461 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
    11521 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
    11581 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
    11641 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
    11701 taagctgatc gccagaaaga aggactggga ccctaagaag t
//
