LOCUS       sequence-338139-       11742 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 11742)
  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 11742)
  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..11742
                     /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"
     intron          1053..1280
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1292..1301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1301..1366
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1373..1393
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1418..5518
                     /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             1418..5518
                     /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             7691..7738
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7754..7807
                     /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             7808..8407
                     /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(7808..7827)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8304..8324
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(8435..8452)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8441..8648
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8657..8673
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8657..8673
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        complement(8716..8791)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(8809..9057)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     repeat_region   9066..9206
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   9066..9195
                     /label=AAV2 ITR
     rep_origin      9281..9736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9368..9387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9578..9599
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9753..9772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9872..9894
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9932..9950)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10018..10122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10123..10191,10192..10983)
                     /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(10341..10360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11154..11742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11643..11662
                     /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 gggnnnsgsg
      901 sgcsrggsgg ggsggggsgg ggsgrggggs ggggsggggs grggsggrgr ggkgsggcgg
      961 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
     1021 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
     1081 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
     1141 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt
     1201 aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc
     1261 ctgaaatcac tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg
     1321 agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa
     1381 gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct
     1441 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     1501 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     1561 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     1621 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     1681 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     1741 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     1801 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     1861 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     1921 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     1981 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     2041 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     2101 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     2161 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     2221 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     2281 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     2341 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     2401 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     2461 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     2521 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     2581 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     2641 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     2701 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     2761 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     2821 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     2881 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     2941 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     3001 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     3061 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     3121 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     3181 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     3241 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     3301 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     3361 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     3421 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     3481 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     3541 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     3601 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     3661 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     3721 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     3781 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     3841 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     3901 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     3961 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     4021 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     4081 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     4141 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     4201 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     4261 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     4321 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     4381 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     4441 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     4501 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     4561 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     4621 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     4681 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     4741 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     4801 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     4861 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     4921 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     4981 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     5041 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     5101 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     5161 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     5221 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     5281 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     5341 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     5401 caccatcgac cggaagaggt acaccagcac gaaagaggtg ctggacgcca ccctgatcca
     5461 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggtgactc
     5521 tggaggatct agcggaggat cctctggcag cgagacacca ggaacaagcg agtcagcaac
     5581 accagagagc agtggcggca gcagcggcgg cagcagcacc ctaaatatag aagatgagta
     5641 tcggctacat gagacctcaa aagagccaga tgtttctcta gggtccacat ggctgtctga
     5701 ttttcctcag gcctgggcgg aaaccggggg catgggactg gcagttcgcc aagctcctct
     5761 gatcatacct ctgaaagcaa cctctacccc cgtgtccata aaacaatacc ccatgtcaca
     5821 agaagccaga ctggggatca agccccacat acagagactg ttggaccagg gaatactggt
     5881 accctgccag tccccctgga acacgcccct gctacccgtt aagaaaccag ggactaatga
     5941 ttataggcct gtccaggatc tgagagaagt caacaagcgg gtggaggaca tccaccccac
     6001 cgtgcccaac ccttacaacc tcttgagcgg gctcccaccg tcccaccagt ggtacactgt
     6061 gcttgattta aaggatgcct ttttctgcct gagactccac cccaccagtc agcctctctt
     6121 cgcctttgag tggagagatc cagagatggg aatctcagga caattgacct ggaccagact
     6181 cccacagggt ttcaaaaaca gtcccaccct gtttaatgag gcactgcaca gagacctagc
     6241 agacttccgg atccagcacc cagacttgat cctgctacag tacgtggatg acttactgct
     6301 ggccgccact tctgagctag actgccaaca aggtactcgg gccctgttac aaaccctagg
     6361 gaacctcggg tatcgggcct cggccaagaa agcccaaatt tgccagaaac aggtcaagta
     6421 tctggggtat cttctaaaag agggtcagag atggctgact gaggccagaa aagagactgt
     6481 gatggggcag cctactccaa agacccctcg acaactaagg gagttcctag ggaaggcagg
     6541 cttctgtcgc ctcttcatcc ctgggtttgc agaaatggca gcccccctgt accctctcac
     6601 caaaccgggg actctgttta attggggccc agaccaacaa aaggcctatc aagaaatcaa
     6661 gcaagccctt ctaactgccc cagccctggg gttgccagat ttgactaagc cctttgaact
     6721 ctttgtcgac gagaagcagg gctacgccaa aggtgtccta acgcaaaaac tgggaccttg
     6781 gcgtcggccg gtggcctacc tgtccaaaaa gctagaccca gtagcagctg ggtggccccc
     6841 ttgcctacgg atggtagcag ccattgccgt actgacaaag gatgcaggca agctaaccat
     6901 gggacagcca ctagtcattc tggcccccca tgcagtagag gcactagtca aacaaccccc
     6961 cgaccgctgg ctttccaacg cccggatgac tcactatcag gccttgcttt tggacacgga
     7021 ccgggtccag ttcggaccgg tggtagccct gaacccggct acgctgctcc cactgcctga
     7081 ggaagggctg caacacaact gccttgatat cctggccgaa gcccacggaa cccgacccga
     7141 cctaacggac cagccgctcc cagacgccga ccacacctgg tacacggatg gaagcagtct
     7201 cttacaagag ggacagcgta aggcgggagc tgcggtgacc accgagaccg aggtaatctg
     7261 ggctaaagcc ctgccagccg ggacatccgc tcagcgggct gaactgatag cactcaccca
     7321 ggccctaaag atggcagaag gtaagaagct aaatgtttat actgatagcc gttatgcttt
     7381 tgctactgcc catatccatg gagaaatata cagaaggcgt gggtggctca catcagaagg
     7441 caaagagatc aaaaataaag acgagatctt ggccctacta aaagccctct ttctgcccaa
     7501 aagacttagc ataatccatt gtccaggaca tcaaaaggga cacagcgccg aggctagagg
     7561 caaccggatg gctgaccaag cggcccgaaa ggcagccatc acagagactc cagacacctc
     7621 taccctcctc atagaaaatt catcaccctc tggcggctca aaaagaaccg ccgacggcag
     7681 cgaattcgag aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagga
     7741 attcggcagt ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc
     7801 tggcccaatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     7861 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     7921 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     7981 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     8041 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     8101 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     8161 caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg
     8221 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     8281 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     8341 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     8401 tgcctgagaa ttctaactag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     8461 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     8521 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     8581 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat
     8641 gctggggagc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatacgaa
     8701 tcctgcagga aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct
     8761 tattttaact tgctatttct agctctaaaa cgtcttctcg aagaccccgg tgtttcgtcc
     8821 tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg taagcatatg
     8881 atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt attactttct
     8941 acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc caattatctc
     9001 tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata ggccctcgcg
     9061 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     9121 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     9181 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     9241 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     9301 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     9361 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     9421 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     9481 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     9541 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     9601 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     9661 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     9721 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     9781 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     9841 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     9901 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     9961 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
    10021 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
    10081 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
    10141 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
    10201 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
    10261 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
    10321 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
    10381 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
    10441 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
    10501 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
    10561 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
    10621 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
    10681 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
    10741 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
    10801 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca
    10861 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
    10921 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
    10981 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    11041 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    11101 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
    11161 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
    11221 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
    11281 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    11341 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    11401 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    11461 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    11521 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
    11581 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    11641 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    11701 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aa
//
