LOCUS       sequence-341857-       11756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Delivers all prime editing nuclease components targeting the HEK3 
            gene for a CTT inserrtion, in a single, puromycin selectable 
            plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11756)
  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 11756)
  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-Nuclease-Puro HEK3 CTT ins
FEATURES             Location/Qualifiers
     source          1..11756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..310
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        331..406
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        499..784
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1063..1290
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1302..1311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1311..1376
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1383..1403
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1428..5528
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1428..5528
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7701..7748
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7764..7817
                     /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             7818..8417
                     /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(7818..7837)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8314..8334
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(8445..8462)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8451..8658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8667..8683
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8667..8683
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        complement(8726..8801)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(8823..9071)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     repeat_region   9080..9220
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   9080..9209
                     /label=AAV2 ITR
     rep_origin      9295..9750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9382..9401)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9592..9613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9767..9786)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9886..9908
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9946..9964)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10032..10136
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10137..10205,10206..10997)
                     /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(10355..10374)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11168..11756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11657..11676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      121 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      181 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      241 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      301 acgaaacacc ggcccagact gagcacgtga gttttagagc tagaaatagc aagttaaaat
      361 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctctg ccatcaaagc
      421 gtgctcagtc tgtttttttg ttctagtccg tttttagcgc gtgcgccaat tctgcagaca
      481 aatggctcta gaggtacccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      541 caacgacccc cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca
      601 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      661 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta
      721 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      781 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      841 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      901 gggnnnsgsg cgccrggsgg ggsggggsgg ggsgrggggs ggggsggggs grggsggrgr
      961 ggkgsggsgg crgccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
     1021 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
     1081 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
     1141 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg
     1201 agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg
     1261 gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg gactataagg
     1321 accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg
     1381 ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca
     1441 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     1501 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     1561 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     1621 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     1681 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     1741 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     1801 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     1861 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     1921 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     1981 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     2041 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     2101 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     2161 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     2221 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     2281 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     2341 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     2401 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     2461 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     2521 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     2581 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     2641 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     2701 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     2761 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     2821 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     2881 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     2941 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     3001 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     3061 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     3121 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     3181 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     3241 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     3301 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     3361 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     3421 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     3481 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     3541 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     3601 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     3661 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     3721 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     3781 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     3841 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     3901 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     3961 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     4021 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     4081 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     4141 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     4201 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     4261 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     4321 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     4381 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     4441 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     4501 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     4561 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     4621 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     4681 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     4741 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     4801 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     4861 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     4921 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     4981 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     5041 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     5101 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     5161 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     5221 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     5281 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     5341 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     5401 actttgacac caccatcgac cggaagaggt acaccagcac gaaagaggtg ctggacgcca
     5461 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     5521 gaggtgactc tggaggatct agcggaggat cctctggcag cgagacacca ggaacaagcg
     5581 agtcagcaac accagagagc agtggcggca gcagcggcgg cagcagcacc ctaaatatag
     5641 aagatgagta tcggctacat gagacctcaa aagagccaga tgtttctcta gggtccacat
     5701 ggctgtctga ttttcctcag gcctgggcgg aaaccggggg catgggactg gcagttcgcc
     5761 aagctcctct gatcatacct ctgaaagcaa cctctacccc cgtgtccata aaacaatacc
     5821 ccatgtcaca agaagccaga ctggggatca agccccacat acagagactg ttggaccagg
     5881 gaatactggt accctgccag tccccctgga acacgcccct gctacccgtt aagaaaccag
     5941 ggactaatga ttataggcct gtccaggatc tgagagaagt caacaagcgg gtggaggaca
     6001 tccaccccac cgtgcccaac ccttacaacc tcttgagcgg gctcccaccg tcccaccagt
     6061 ggtacactgt gcttgattta aaggatgcct ttttctgcct gagactccac cccaccagtc
     6121 agcctctctt cgcctttgag tggagagatc cagagatggg aatctcagga caattgacct
     6181 ggaccagact cccacagggt ttcaaaaaca gtcccaccct gtttaatgag gcactgcaca
     6241 gagacctagc agacttccgg atccagcacc cagacttgat cctgctacag tacgtggatg
     6301 acttactgct ggccgccact tctgagctag actgccaaca aggtactcgg gccctgttac
     6361 aaaccctagg gaacctcggg tatcgggcct cggccaagaa agcccaaatt tgccagaaac
     6421 aggtcaagta tctggggtat cttctaaaag agggtcagag atggctgact gaggccagaa
     6481 aagagactgt gatggggcag cctactccaa agacccctcg acaactaagg gagttcctag
     6541 ggaaggcagg cttctgtcgc ctcttcatcc ctgggtttgc agaaatggca gcccccctgt
     6601 accctctcac caaaccgggg actctgttta attggggccc agaccaacaa aaggcctatc
     6661 aagaaatcaa gcaagccctt ctaactgccc cagccctggg gttgccagat ttgactaagc
     6721 cctttgaact ctttgtcgac gagaagcagg gctacgccaa aggtgtccta acgcaaaaac
     6781 tgggaccttg gcgtcggccg gtggcctacc tgtccaaaaa gctagaccca gtagcagctg
     6841 ggtggccccc ttgcctacgg atggtagcag ccattgccgt actgacaaag gatgcaggca
     6901 agctaaccat gggacagcca ctagtcattc tggcccccca tgcagtagag gcactagtca
     6961 aacaaccccc cgaccgctgg ctttccaacg cccggatgac tcactatcag gccttgcttt
     7021 tggacacgga ccgggtccag ttcggaccgg tggtagccct gaacccggct acgctgctcc
     7081 cactgcctga ggaagggctg caacacaact gccttgatat cctggccgaa gcccacggaa
     7141 cccgacccga cctaacggac cagccgctcc cagacgccga ccacacctgg tacacggatg
     7201 gaagcagtct cttacaagag ggacagcgta aggcgggagc tgcggtgacc accgagaccg
     7261 aggtaatctg ggctaaagcc ctgccagccg ggacatccgc tcagcgggct gaactgatag
     7321 cactcaccca ggccctaaag atggcagaag gtaagaagct aaatgtttat actgatagcc
     7381 gttatgcttt tgctactgcc catatccatg gagaaatata cagaaggcgt gggtggctca
     7441 catcagaagg caaagagatc aaaaataaag acgagatctt ggccctacta aaagccctct
     7501 ttctgcccaa aagacttagc ataatccatt gtccaggaca tcaaaaggga cacagcgccg
     7561 aggctagagg caaccggatg gctgaccaag cggcccgaaa ggcagccatc acagagactc
     7621 cagacacctc taccctcctc atagaaaatt catcaccctc tggcggctca aaaagaaccg
     7681 ccgacggcag cgaattcgag aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
     7741 agaaaaagga attcggcagt ggagagggca gaggaagtct gctaacatgc ggtgacgtcg
     7801 aggagaatcc tggcccaatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     7861 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     7921 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     7981 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     8041 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     8101 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     8161 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
     8221 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     8281 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     8341 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     8401 gcaagcccgg tgcctgagaa ttctaactag agctcgctga tcagcctcga ctgtgccttc
     8461 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     8521 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     8581 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa
     8641 tagcaggcat gctggggagc ggccgctcta gaactagtgg atcccccggg ctgcaggaat
     8701 tcgatacgaa tcctgcagga aaaaagcacc gactcggtgc cactttttca agttgataac
     8761 ggactagcct tattttaact tgctatttct agctctaaaa caatcttgca gtgttgcctg
     8821 acggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata ctttcaagtt
     8881 acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa ctacccaaga
     8941 aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac agtcaaatta
     9001 attccaatta tctctctaac agccttgtat cgtatatgca aatatgaagg aatcatggga
     9061 aataggccct cgcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     9121 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     9181 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     9241 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     9301 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     9361 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     9421 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     9481 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     9541 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     9601 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     9661 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     9721 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     9781 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     9841 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     9901 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     9961 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
    10021 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
    10081 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
    10141 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
    10201 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
    10261 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
    10321 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
    10381 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
    10441 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
    10501 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
    10561 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
    10621 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
    10681 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
    10741 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
    10801 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg
    10861 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
    10921 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
    10981 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
    11041 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
    11101 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
    11161 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
    11221 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
    11281 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
    11341 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
    11401 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
    11461 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
    11521 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
    11581 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
    11641 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
    11701 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaa
//
