LOCUS       sequence-348843-        9121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expressing a fusion protein of spCas9 and the 3' 
            exonuclease domain of POLI, and sgRNA targeting human CLCN5 gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9121)
  AUTHORS   Yoo KW, Yadav MK, Song Q, Atala A, Lu B
  TITLE     Targeting DNA polymerase to DNA double-strand breaks reduces DNA 
            deletion size and increases templated insertions generated by 
            CRISPR/Cas9.
  JOURNAL   Nucleic Acids Res. 2022 Mar 22. pii: 6552087. doi: 
            10.1093/nar/gkac186.
  PUBMED    35323942
REFERENCE   2  (bases 1 to 9121)
  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. 2022 Mar 22. pii: 6552087. doi: 10.1093/nar/gkac186."
COMMENT     Sequence Label: pspCas9-3exo-Tetra-com-CLCN5-sp-g1
FEATURES             Location/Qualifiers
     source          1..9121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..291
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          571..798
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      810..819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             819..884
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             891..911
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             936..5036
                     /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             936..5036
                     /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             5037..5084
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5778..5795)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5784..5991
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6003..6143
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6003..6132
                     /label=AAV2 ITR
     rep_origin      6218..6673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6305..6324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6515..6536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6690..6709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6809..6831
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6869..6887)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6955..7059
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7060..7128,7129..7920)
                     /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(7278..7297)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8091..8679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8580..8599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8741..8989
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8741..8761
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8912..8931
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
       61 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      121 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      181 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
      241 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
      301 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
      361 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg
      421 ccrggsgggg sggggsgggg sgrggggsgg ggsggggsgr ggsggrgagg kgcggcggca
      481 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
      541 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
      601 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      661 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa
      721 gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct
      781 gaaatcactt tttttcaggt tggaccggtg ccaccatgga ctataaggac cacgacggag
      841 actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga
      901 agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg
      961 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     1021 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     1081 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     1141 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     1201 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     1261 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     1321 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     1381 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     1441 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     1501 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     1561 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     1621 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     1681 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     1741 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     1801 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     1861 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     1921 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     1981 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     2041 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     2101 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     2161 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     2221 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     2281 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     2341 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     2401 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     2461 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     2521 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     2581 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     2641 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     2701 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     2761 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     2821 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     2881 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     2941 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     3001 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     3061 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     3121 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     3181 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     3241 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     3301 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     3361 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     3421 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     3481 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     3541 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     3601 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     3661 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     3721 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     3781 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     3841 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     3901 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     3961 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     4021 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     4081 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     4141 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     4201 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     4261 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     4321 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     4381 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     4441 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     4501 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     4561 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     4621 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     4681 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     4741 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     4801 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     4861 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     4921 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     4981 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa
     5041 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagggatcc tctgagacac
     5101 caggcacctc cgagtctgcc acccctgagg gaggagcacc agaggtgaca gccaccgtga
     5161 tctcctacga taactatgtg acaatcctgg acgaggagac actgaaggcc tggatcgcca
     5221 agctggagaa ggcccccgtg ttcgcctttg atacagagac agacagcctg gataacatct
     5281 ccgccaatct ggtgggcctg tctttcgcca tcgagcctgg cgtggccgcc tatatcccag
     5341 tggcccacga ctacctggat gcccccgacc agatcagcag ggagagagcc ctggagctgc
     5401 tgaagcctct gctggaggat gagaaggccc tgaaggtcgg ccagaacctg aagtatgaca
     5461 ggggcatcct ggccaattac ggcatcgagc tgagaggcat cgcctttgac accatgctgg
     5521 agtcttatat cctgaatagc gtggcaggcc ggcacgacat ggattccctg gccgagaggt
     5581 ggctgaagca caagacaatc accttcgagg agatcgccgg caagggcaag aaccagctga
     5641 ccttcaacca gatcgccctg gaggaggcag gcaggtacgc agcagaggac gcagatgtga
     5701 ccctgcagct gcacctgaag atgtggccag atctgcagaa gcactagtct agaaaattcc
     5761 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     5821 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     5881 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     5941 gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg aggtacggcc
     6001 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     6061 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     6121 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     6181 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     6241 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     6301 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6361 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6421 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6481 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6541 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     6601 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     6661 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6721 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6781 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6841 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     6901 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6961 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7021 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7081 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7141 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7201 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7261 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7321 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7381 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7441 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7501 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7561 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7621 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7681 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7741 tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg
     7801 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7861 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7921 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7981 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     8041 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     8101 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     8161 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     8221 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     8281 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8341 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8401 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8461 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8521 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8581 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8641 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8701 ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat ttcccatgat
     8761 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
     8821 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     8881 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     8941 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg gaggacaagt
     9001 cgtacaatgg gtttgagagc taggccctga atgcctgcga gcatcccacg gcctagcaag
     9061 ttcaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
     9121 g
//
