LOCUS       sequence-339091-       11753 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 11753)
  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 11753)
  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..11753
                     /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"
     promoter        786..1059
                     /label=chicken beta-actin promoter
     intron          1060..1287
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1299..1308
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1308..1373
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1380..1400
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1425..5525
                     /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             1425..5525
                     /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             7698..7745
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7761..7814
                     /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             7815..8414
                     /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(7815..7834)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8311..8331
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(8442..8459)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8448..8655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8664..8680
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8664..8680
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        complement(8723..8798)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(8820..9068)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     repeat_region   9077..9217
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   9077..9206
                     /label=AAV2 ITR
     rep_origin      9292..9747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9379..9398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9589..9610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9764..9783)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9883..9905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9943..9961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10029..10133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10134..10202,10203..10994)
                     /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(10352..10371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11165..11753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11654..11673
                     /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 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
      961 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     1021 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
     1081 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
     1141 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc
     1201 aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag
     1261 cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatggac tataaggacc
     1321 acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc
     1381 caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca
     1441 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     1501 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     1561 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     1621 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     1681 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     1741 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     1801 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     1861 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     1921 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     1981 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     2041 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     2101 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
     2161 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     2221 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     2281 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     2341 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     2401 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     2461 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     2521 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     2581 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     2641 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     2701 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     2761 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     2821 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     2881 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     2941 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     3001 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     3061 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     3121 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     3181 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     3241 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     3301 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     3361 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     3421 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     3481 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     3541 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     3601 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     3661 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     3721 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     3781 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     3841 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     3901 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     3961 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     4021 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     4081 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     4141 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     4201 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     4261 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     4321 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     4381 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     4441 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     4501 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     4561 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     4621 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     4681 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     4741 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     4801 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     4861 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     4921 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     4981 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     5041 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     5101 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     5161 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     5221 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     5281 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     5341 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     5401 ttgacaccac catcgaccgg aagaggtaca ccagcacgaa agaggtgctg gacgccaccc
     5461 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     5521 gtgactctgg aggatctagc ggaggatcct ctggcagcga gacaccagga acaagcgagt
     5581 cagcaacacc agagagcagt ggcggcagca gcggcggcag cagcacccta aatatagaag
     5641 atgagtatcg gctacatgag acctcaaaag agccagatgt ttctctaggg tccacatggc
     5701 tgtctgattt tcctcaggcc tgggcggaaa ccgggggcat gggactggca gttcgccaag
     5761 ctcctctgat catacctctg aaagcaacct ctacccccgt gtccataaaa caatacccca
     5821 tgtcacaaga agccagactg gggatcaagc cccacataca gagactgttg gaccagggaa
     5881 tactggtacc ctgccagtcc ccctggaaca cgcccctgct acccgttaag aaaccaggga
     5941 ctaatgatta taggcctgtc caggatctga gagaagtcaa caagcgggtg gaggacatcc
     6001 accccaccgt gcccaaccct tacaacctct tgagcgggct cccaccgtcc caccagtggt
     6061 acactgtgct tgatttaaag gatgcctttt tctgcctgag actccacccc accagtcagc
     6121 ctctcttcgc ctttgagtgg agagatccag agatgggaat ctcaggacaa ttgacctgga
     6181 ccagactccc acagggtttc aaaaacagtc ccaccctgtt taatgaggca ctgcacagag
     6241 acctagcaga cttccggatc cagcacccag acttgatcct gctacagtac gtggatgact
     6301 tactgctggc cgccacttct gagctagact gccaacaagg tactcgggcc ctgttacaaa
     6361 ccctagggaa cctcgggtat cgggcctcgg ccaagaaagc ccaaatttgc cagaaacagg
     6421 tcaagtatct ggggtatctt ctaaaagagg gtcagagatg gctgactgag gccagaaaag
     6481 agactgtgat ggggcagcct actccaaaga cccctcgaca actaagggag ttcctaggga
     6541 aggcaggctt ctgtcgcctc ttcatccctg ggtttgcaga aatggcagcc cccctgtacc
     6601 ctctcaccaa accggggact ctgtttaatt ggggcccaga ccaacaaaag gcctatcaag
     6661 aaatcaagca agcccttcta actgccccag ccctggggtt gccagatttg actaagccct
     6721 ttgaactctt tgtcgacgag aagcagggct acgccaaagg tgtcctaacg caaaaactgg
     6781 gaccttggcg tcggccggtg gcctacctgt ccaaaaagct agacccagta gcagctgggt
     6841 ggcccccttg cctacggatg gtagcagcca ttgccgtact gacaaaggat gcaggcaagc
     6901 taaccatggg acagccacta gtcattctgg ccccccatgc agtagaggca ctagtcaaac
     6961 aaccccccga ccgctggctt tccaacgccc ggatgactca ctatcaggcc ttgcttttgg
     7021 acacggaccg ggtccagttc ggaccggtgg tagccctgaa cccggctacg ctgctcccac
     7081 tgcctgagga agggctgcaa cacaactgcc ttgatatcct ggccgaagcc cacggaaccc
     7141 gacccgacct aacggaccag ccgctcccag acgccgacca cacctggtac acggatggaa
     7201 gcagtctctt acaagaggga cagcgtaagg cgggagctgc ggtgaccacc gagaccgagg
     7261 taatctgggc taaagccctg ccagccggga catccgctca gcgggctgaa ctgatagcac
     7321 tcacccaggc cctaaagatg gcagaaggta agaagctaaa tgtttatact gatagccgtt
     7381 atgcttttgc tactgcccat atccatggag aaatatacag aaggcgtggg tggctcacat
     7441 cagaaggcaa agagatcaaa aataaagacg agatcttggc cctactaaaa gccctctttc
     7501 tgcccaaaag acttagcata atccattgtc caggacatca aaagggacac agcgccgagg
     7561 ctagaggcaa ccggatggct gaccaagcgg cccgaaaggc agccatcaca gagactccag
     7621 acacctctac cctcctcata gaaaattcat caccctctgg cggctcaaaa agaaccgccg
     7681 acggcagcga attcgagaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
     7741 aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     7801 agaatcctgg cccaatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     7861 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     7921 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     7981 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     8041 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     8101 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     8161 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg
     8221 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     8281 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     8341 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     8401 agcccggtgc ctgagaattc taactagagc tcgctgatca gcctcgactg tgccttctag
     8461 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     8521 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     8581 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag
     8641 caggcatgct ggggagcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg
     8701 atacgaatcc tgcaggaaaa aagcaccgac tcggtgccac tttttcaagt tgataacgga
     8761 ctagccttat tttaacttgc tatttctagc tctaaaacaa tcttgcagtg ttgcctgacg
     8821 gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg
     8881 gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta cccaagaaat
     8941 tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt caaattaatt
     9001 ccaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat catgggaaat
     9061 aggccctcgc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     9121 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     9181 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     9241 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     9301 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     9361 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     9421 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     9481 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     9541 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     9601 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     9661 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     9721 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     9781 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     9841 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     9901 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     9961 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
    10021 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
    10081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
    10141 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
    10201 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
    10261 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
    10321 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
    10381 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
    10441 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
    10501 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
    10561 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
    10621 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
    10681 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
    10741 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
    10801 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
    10861 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
    10921 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
    10981 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
    11041 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
    11101 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
    11161 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
    11221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
    11281 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
    11341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
    11401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
    11461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
    11521 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
    11581 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
    11641 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
    11701 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaa
//
