LOCUS       sequence-339088-       11739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Delivers all prime editing nuclease components in a single, 
            puromycin selectable plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11739)
  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 11739)
  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
FEATURES             Location/Qualifiers
     source          1..11739
                     /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        329..404
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        489..774
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        776..1049
                     /label=chicken beta-actin promoter
     intron          1050..1277
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1289..1298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1298..1363
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1370..1390
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1415..5515
                     /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             1415..5515
                     /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             7688..7735
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7751..7804
                     /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             7805..8404
                     /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(7805..7824)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8301..8321
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(8432..8449)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8438..8645
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8654..8670
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8654..8670
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        complement(8713..8788)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(8806..9054)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     repeat_region   9063..9203
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   9063..9192
                     /label=AAV2 ITR
     rep_origin      9278..9733
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9365..9384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9575..9596
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9750..9769)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9869..9891
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9929..9947)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10015..10119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10120..10188,10189..10980)
                     /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(10338..10357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11151..11739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11640..11659
                     /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 gggtcttcga gaagacctgt tttagagcta gaaatagcaa gttaaaataa
      361 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgcgggtct tcgagaagac
      421 cttttttttg ttctagtccg tttttagcgc gtgcgccaat tctgcagaca aatggctcta
      481 gaggtacccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      541 cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag
      601 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      661 cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta
      721 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
      781 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
      841 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggcgcgcgc
      901 caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag
      961 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
     1021 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc
     1081 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
     1141 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag
     1201 ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg
     1261 aaatcacttt ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga
     1321 ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa
     1381 gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga
     1441 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     1501 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     1561 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     1621 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     1681 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     1741 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     1801 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     1861 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     1921 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     1981 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     2041 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     2101 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     2161 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     2221 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     2281 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     2341 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     2401 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     2461 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     2521 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     2581 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     2641 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     2701 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     2761 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     2821 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     2881 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     2941 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     3001 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     3061 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     3121 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     3181 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     3241 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     3301 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     3361 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     3421 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     3481 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     3541 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     3601 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     3661 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     3721 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     3781 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     3841 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     3901 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     3961 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     4021 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     4081 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     4141 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     4201 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     4261 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     4321 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     4381 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     4441 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     4501 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     4561 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     4621 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     4681 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     4741 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     4801 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     4861 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     4921 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     4981 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     5041 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     5101 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     5161 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     5221 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     5281 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     5341 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     5401 catcgaccgg aagaggtaca ccagcacgaa agaggtgctg gacgccaccc tgatccacca
     5461 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg
     5521 aggatctagc ggaggatcct ctggcagcga gacaccagga acaagcgagt cagcaacacc
     5581 agagagcagt ggcggcagca gcggcggcag cagcacccta aatatagaag atgagtatcg
     5641 gctacatgag acctcaaaag agccagatgt ttctctaggg tccacatggc tgtctgattt
     5701 tcctcaggcc tgggcggaaa ccgggggcat gggactggca gttcgccaag ctcctctgat
     5761 catacctctg aaagcaacct ctacccccgt gtccataaaa caatacccca tgtcacaaga
     5821 agccagactg gggatcaagc cccacataca gagactgttg gaccagggaa tactggtacc
     5881 ctgccagtcc ccctggaaca cgcccctgct acccgttaag aaaccaggga ctaatgatta
     5941 taggcctgtc caggatctga gagaagtcaa caagcgggtg gaggacatcc accccaccgt
     6001 gcccaaccct tacaacctct tgagcgggct cccaccgtcc caccagtggt acactgtgct
     6061 tgatttaaag gatgcctttt tctgcctgag actccacccc accagtcagc ctctcttcgc
     6121 ctttgagtgg agagatccag agatgggaat ctcaggacaa ttgacctgga ccagactccc
     6181 acagggtttc aaaaacagtc ccaccctgtt taatgaggca ctgcacagag acctagcaga
     6241 cttccggatc cagcacccag acttgatcct gctacagtac gtggatgact tactgctggc
     6301 cgccacttct gagctagact gccaacaagg tactcgggcc ctgttacaaa ccctagggaa
     6361 cctcgggtat cgggcctcgg ccaagaaagc ccaaatttgc cagaaacagg tcaagtatct
     6421 ggggtatctt ctaaaagagg gtcagagatg gctgactgag gccagaaaag agactgtgat
     6481 ggggcagcct actccaaaga cccctcgaca actaagggag ttcctaggga aggcaggctt
     6541 ctgtcgcctc ttcatccctg ggtttgcaga aatggcagcc cccctgtacc ctctcaccaa
     6601 accggggact ctgtttaatt ggggcccaga ccaacaaaag gcctatcaag aaatcaagca
     6661 agcccttcta actgccccag ccctggggtt gccagatttg actaagccct ttgaactctt
     6721 tgtcgacgag aagcagggct acgccaaagg tgtcctaacg caaaaactgg gaccttggcg
     6781 tcggccggtg gcctacctgt ccaaaaagct agacccagta gcagctgggt ggcccccttg
     6841 cctacggatg gtagcagcca ttgccgtact gacaaaggat gcaggcaagc taaccatggg
     6901 acagccacta gtcattctgg ccccccatgc agtagaggca ctagtcaaac aaccccccga
     6961 ccgctggctt tccaacgccc ggatgactca ctatcaggcc ttgcttttgg acacggaccg
     7021 ggtccagttc ggaccggtgg tagccctgaa cccggctacg ctgctcccac tgcctgagga
     7081 agggctgcaa cacaactgcc ttgatatcct ggccgaagcc cacggaaccc gacccgacct
     7141 aacggaccag ccgctcccag acgccgacca cacctggtac acggatggaa gcagtctctt
     7201 acaagaggga cagcgtaagg cgggagctgc ggtgaccacc gagaccgagg taatctgggc
     7261 taaagccctg ccagccggga catccgctca gcgggctgaa ctgatagcac tcacccaggc
     7321 cctaaagatg gcagaaggta agaagctaaa tgtttatact gatagccgtt atgcttttgc
     7381 tactgcccat atccatggag aaatatacag aaggcgtggg tggctcacat cagaaggcaa
     7441 agagatcaaa aataaagacg agatcttggc cctactaaaa gccctctttc tgcccaaaag
     7501 acttagcata atccattgtc caggacatca aaagggacac agcgccgagg ctagaggcaa
     7561 ccggatggct gaccaagcgg cccgaaaggc agccatcaca gagactccag acacctctac
     7621 cctcctcata gaaaattcat caccctctgg cggctcaaaa agaaccgccg acggcagcga
     7681 attcgagaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggaatt
     7741 cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     7801 cccaatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     7861 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     7921 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     7981 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     8041 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     8101 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     8161 ggagcccgcg tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct
     8221 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     8281 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     8341 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     8401 ctgagaattc taactagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     8461 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     8521 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     8581 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     8641 ggggagcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg atacgaatcc
     8701 tgcaggaaaa aagcaccgac tcggtgccac tttttcaagt tgataacgga ctagccttat
     8761 tttaacttgc tatttctagc tctaaaacgt cttctcgaag accccggtgt ttcgtccttt
     8821 ccacaagata tataaagcca agaaatcgaa atactttcaa gttacggtaa gcatatgata
     8881 gtccatttta aaacataatt ttaaaactgc aaactaccca agaaattatt actttctacg
     8941 tcacgtattt tgtactaata tctttgtgtt tacagtcaaa ttaattccaa ttatctctct
     9001 aacagccttg tatcgtatat gcaaatatga aggaatcatg ggaaataggc cctcgcggcc
     9061 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     9121 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     9181 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     9241 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     9301 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     9361 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     9421 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     9481 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     9541 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     9601 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     9661 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     9721 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     9781 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     9841 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     9901 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     9961 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
    10021 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
    10081 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
    10141 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
    10201 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
    10261 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
    10321 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
    10381 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
    10441 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
    10501 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
    10561 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
    10621 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
    10681 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
    10741 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
    10801 tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg
    10861 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
    10921 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
    10981 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
    11041 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
    11101 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
    11161 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
    11221 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
    11281 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
    11341 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
    11401 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
    11461 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
    11521 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
    11581 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
    11641 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
    11701 tttttgtgat gctcgtcagg ggggcggagc ctatggaaa
//
