LOCUS       sequence-373356-       10635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sgRNA targeting Dora gene and Cas9-Puro in Drosophila S2 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10635)
  AUTHORS   Kingston ER, Blodgett LW, Bartel DP
  TITLE     Endogenous transcripts direct microRNA degradation in Drosophila, 
            and this targeted degradation is required for proper embryonic 
            development.
  JOURNAL   Mol Cell. 2022 Sep 15. pii: S1097-2765(22)00849-8. doi: 
            10.1016/j.molcel.2022.08.029.
  PUBMED    36150386
REFERENCE   2  (bases 1 to 10635)
  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: "Mol Cell. 
            2022 Sep 15. pii: S1097-2765(22)00849-8. doi: 
            10.1016/j.molcel.2022.08.029."
COMMENT     Sequence Label: pAc-sgRNA-Cas9.dora_3
FEATURES             Location/Qualifiers
     source          1..10635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        790..1189
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        1211..1286
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1438..1456)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1524..1628
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1629..1697,1698..2489)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1847..1866)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2660..3248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3149..3168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3849..3983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3886..3905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3940..3959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3991..4590)
                     /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(4074..4094)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4571..4590
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4615..4668)
                     /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             complement(4684..4731)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4732..8832)
                     /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             complement(4732..8832)
                     /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             complement(8857..8877)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8884..8949)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 ccttccacta ccgttttgcc gcatttggta ttgccatctc gcttttttgg ctttactttg
       61 ctttttgaat aagccaagaa actgaaatca aagcggacag tccgaaaatt gaatattcca
      121 aaaaacaaca attgcacatg cactgcgttc ggcaaaagac gcagaaattc aagccttttg
      181 gtctggcatc atcttaaaat atcatcagta tcatcattgc cattataaat acataagaaa
      241 atgagaatac attaaagata tacatacaat ctttgtcata gtcaacatat tttgcaacaa
      301 aattatctat agataacatg tttttgtaga aaattccact aactgatttt cgtataaata
      361 caaattattc tcattcttca aaatactcgc cgacgtcatc gcgaaattcc tcagcattct
      421 tcgacaaatg aagagagaca gcgagtgaca caatcttatc ttgccgacga gtctttgaaa
      481 atggccccgg gtaacggtga aaaacatata gccaagaatg gagcgatcgc caggcgatgg
      541 aaattggctt caaattgctc cgaatatggc cagttggtca gttggcattg gattgtggag
      601 agatactaga tcagctcgtt ctccttctcc tccgcattct cctactccgc ccgaagacgc
      661 ttctttgatc gcttctttct cgaatgtgta catattcatt atgtatttgg tgtttggcga
      721 cgatcagcca gctttaatgg ccataaaacg tgtcttggtt tgtgggccac ataacaggca
      781 gcgagatctg ttcgacttgc agcctgaaat acggcacgag taggaaaagc cgagtcaaat
      841 gccgaatgca gagtctcatt acagcacaat caactcaaga aaaactcgac acttttttac
      901 catttgcact taaatccttt tttattcgtt atgtatactt tttttggtcc ctaaccaaaa
      961 caaaaccaaa ctctcttagt cgtgcctcta tatttaaaac tatcaattta ttatagtcaa
     1021 taaatcgaac tgtgttttca acaaacgaac aataggacac tttgattcta aaggaaattt
     1081 tgaaaatctt aagcagaggg ttcttaagac catttgccaa ttcttataat tctcaactgc
     1141 tctttcctga tgttgatcat ttatataggt atgttttcct caatacttcg catccggctc
     1201 tactcctgtc gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
     1261 ttgaaaaagt ggcaccgagt cggtgctttt ttagatctgg aaaaatgatg tgacagtgga
     1321 aatgagaagt aggtgcatct gcaaagattt gattcagagt tgatgccatt catgattttt
     1381 ttttagaagt atcatttgtg tttttagaat atagaattaa ttcttgaaga cgaaagggcc
     1441 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     1501 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1561 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1621 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1681 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1741 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1801 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1861 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1921 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1981 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2041 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2101 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2161 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2221 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2281 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2341 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2401 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2461 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2521 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2581 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2641 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2701 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2761 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     2821 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2881 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2941 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3001 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3061 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3121 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3181 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3241 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttgaa
     3301 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     3361 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     3421 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     3481 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     3541 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     3601 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     3661 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     3721 ctctcaaggg catcggtcga ctgatcataa tcagccatac cacatttgta gaggttttac
     3781 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3841 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3901 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3961 atgtatctta tcatgtctgg atccctcgag tcaggcaccg ggcttgcggg tcatgcacca
     4021 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     4081 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     4141 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     4201 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     4261 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     4321 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     4381 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     4441 ttgcagctcg gtgacccgct cgatgtggcg gtccgggtcg acggtgtggc gcgtggcggg
     4501 gtagtcggcg aacgcggcgg cgagggtgcg tacggcccgg gggacgtcgt cgcgggtggc
     4561 gaggcgcacc gtgggcttgt actcggtcat gctaccgctg ccgctaccgc tagcagggcc
     4621 gggattctcc tccacgtcac cgcatgttag aagacttcct ctgccctcaa gcttgcctga
     4681 tccctttttc ttttttgcct ggccggcctt tttcgtggcc gccggccttt tgtcgcctcc
     4741 cagctgagac aggtcgatcc gtgtctcgta caggccggtg atgctctggt ggatcagggt
     4801 ggcgtccagc acctctttgg tgctggtgta cctcttccgg tcgatggtgg tgtcaaagta
     4861 cttgaaggcg gcaggggctc ccagattggt cagggtaaac aggtggatga tattctcggc
     4921 ctgctctctg atgggcttat cccggtgctt gttgtaggcg gacagcactt tgtccagatt
     4981 agcgtcggcc aggatcactc tcttggagaa ctcgctgatc tgctcgatga tctcgtccag
     5041 gtagtgcttg tgctgttcca caaacagctg tttctgctca ttatcctcgg gggagccctt
     5101 cagcttctca tagtggctgg ccaggtacag gaagttcaca tatttggagg gcagggccag
     5161 ttcgtttccc ttctgcagtt cgccggcaga ggccagcatt ctcttccggc cgttttccag
     5221 ctcgaacagg gagtacttag gcagcttgat gatcaggtcc tttttcactt ctttgtagcc
     5281 cttggcttcc agaaagtcga tgggattctt ctcgaagctg cttctttcca tgatggtgat
     5341 ccccagcagc tctttcacac tcttcagttt cttggacttg cccttttcca ctttggccac
     5401 caccagcaca gaataggcca cggtggggct gtcgaagccg ccgtacttct tagggtccca
     5461 gtccttcttt ctggcgatca gcttatcgct gttcctcttg ggcaggatag actctttgct
     5521 gaagccgcct gtctgcacct cggtcttttt cacgatattc acttggggca tgctcagcac
     5581 tttccgcacg gtggcaaaat cccggccctt atcccacacg atctccccgg tttcgccgtt
     5641 tgtctcgatc agaggccgct tccggatctc gccgttggcc agggtaatct cggtcttgaa
     5701 aaagttcatg atgttgctgt agaagaagta cttggcggta gccttgccga tttcctgctc
     5761 gctcttggcg atcatcttcc gcacgtcgta caccttgtag tcgccgtaca cgaactcgct
     5821 ttccagctta gggtactttt tgatcagggc ggttcccacg acggcgttca ggtaggcgtc
     5881 gtgggcgtgg tggtagttgt tgatctcgcg cactttgtaa aactggaaat ccttccggaa
     5941 atcggacacc agcttggact tcagggtgat cactttcact tcccggatca gcttgtcatt
     6001 ctcgtcgtac ttagtgttca tccgggagtc caggatctgt gccacgtgct ttgtgatctg
     6061 ccgggtttcc accagctgtc tcttgatgaa gccggcctta tccagttcgc tcaggccgcc
     6121 tctctcggcc ttggtcagat tgtcgaactt tctctgggta atcagcttgg cgttcagcag
     6181 ctgccgccag tagttcttca tcttcttcac gacctcttcg gagggcacgt tgtcgctctt
     6241 gccccggttc ttgtcgcttc tggtcagcac cttgttgtcg atggagtcgt ccttcagaaa
     6301 gctctgaggc acgatatggt ccacatcgta gtcggacagc cggttgatgt ccagttcctg
     6361 gtccacgtac atatcccgcc cattctgcag gtagtacagg tacagcttct cgttctgcag
     6421 ctgggtgttt tccacggggt gttctttcag gatctggctg cccagctctt tgatgccctc
     6481 ttcgatccgc ttcattctct cgcggctgtt cttctgtccc ttctgggtgg tctggttctc
     6541 tctggccatt tcgatcacga tgttctcggg cttgtgccgg cccatcactt tcacgagctc
     6601 gtccaccacc ttcactgtct gcaggatgcc cttcttaatg gcggggctgc cggccagatt
     6661 ggcaatgtgc tcgtgcaggc tatcgccctg gccggacacc tgggctttct ggatgtcctc
     6721 tttaaaggtc aggctgtcgt cgtggatcag ctgcatgaag tttctgttgg cgaagccgtc
     6781 ggacttcagg aaatccagga ttgtcttgcc ggactgcttg tcccggatgc cgttgatcag
     6841 cttccggctc agcctgcccc agccggtgta tctccgccgc ttcagctgct tcatcacttt
     6901 gtcgtcgaac aggtgggcat aggttttcag ccgttcctcg atcatctctc tgtcctcaaa
     6961 cagtgtcagg gtcagcacga tatcttccag aatgtcctcg ttttcctcat tgtccaggaa
     7021 gtccttgtcc ttgataattt tcagcagatc gtggtatgtg cccagggagg cgttgaaccg
     7081 atcttccacg ccggagattt ccacggagtc gaagcactcg attttcttga agtagtcctc
     7141 tttcagctgc ttcacggtca ctttccggtt ggtcttgaac agcaggtcca cgatggcctt
     7201 tttctgctcg ccgctcagga aggcgggctt tctcattccc tcggtcacgt atttcacttt
     7261 ggtcagctcg ttatacacgg tgaagtactc gtacagcagg ctgtgcttgg gcagcacctt
     7321 ctcgttgggc aggttcttat cgaagttggt catccgctcg atgaagctct gggcggaagc
     7381 gcccttgtcc accacttcct cgaagttcca gggggtgatg gtttcctcgc tctttctggt
     7441 catccaggcg aatctgctgt ttcccctggc cagagggccc acgtagtagg ggatgcggaa
     7501 ggtcaggatc ttctcgatct tttcccggtt gtccttcagg aatgggtaaa aatcttcctg
     7561 ccgccgcaga atggcgtgca gctctcccag gtggatctgg tgggggatgc tgccgttgtc
     7621 gaaggtccgc tgcttccgca gcaggtcctc tctgttcagc ttcacgagca gttcctcggt
     7681 gccgtccatc ttttccagga tgggcttgat gaacttgtag aactcttcct ggctggctcc
     7741 gccgtcaatg tagccggcgt agccgttctt gctctggtcg aagaaaatct ctttgtactt
     7801 ctcaggcagc tgctgccgca cgagagcttt cagcagggtc aggtcctggt ggtgctcgtc
     7861 gtatctcttg atcatagagg cgctcagggg ggccttggtg atctcggtgt tcactctcag
     7921 gatgtcgctc agcaggatgg cgtcggacag gttcttggcg gccagaaaca ggtcggcgta
     7981 ctggtcgccg atctgggcca gcaggttgtc caggtcgtcg tcgtaggtgt ccttgctcag
     8041 ctgcagtttg gcatcctcgg ccaggtcgaa gttgctcttg aagttggggg tcaggcccag
     8101 gctcagggca atcaggtttc cgaacaggcc attcttcttc tcgccgggca gctgggcgat
     8161 cagattttcc agccgtctgc tcttgctcag tctggcagac aggatggcct tggcgtccac
     8221 gccgctggcg ttgatggggt tttcctcgaa cagctggttg taggtctgca ccagctggat
     8281 gaacagcttg tccacgtcgc tgttgtcggg gttcaggtcg ccctcgatca ggaagtggcc
     8341 ccggaacttg atcatgtggg ccagggccag atagatcagc cgcaggtcgg ccttgtcggt
     8401 gctgtccacc agtttctttc tcaggtggta gatggtgggg tacttctcgt ggtaggccac
     8461 ctcgtccacg atgttgccga agatggggtg ccgctcgtgc ttcttatcct cttccaccag
     8521 gaaggactct tccagtctgt ggaagaagct gtcgtccacc ttggccatct cgttgctgaa
     8581 gatctcttgc agatagcaga tccggttctt ccgtctggtg tatcttcttc tggcggttct
     8641 cttcagccgg gtggcctcgg ctgtttcgcc gctgtcgaac agcagggctc cgatcaggtt
     8701 cttcttgatg ctgtgccggt cggtgttgcc cagcaccttg aatttcttgc tgggcacctt
     8761 gtactcgtcg gtgatcacgg cccagcccac agagttggtg ccgatgtcca ggccgatgct
     8821 gtacttcttg tcggctgctg ggactccgtg gataccgacc ttccgcttct tctttggggc
     8881 catcttatcg tcatcgtctt tgtaatcaat atcatgatcc ttgtagtctc cgtcgtggtc
     8941 cttatagtcc atgaattcca ccacactgga ctagtaggta ccccgatccg gggtctctgg
     9001 attagacgac tgctggctga tggagcggct ttgtgtcggg aggagtatcc acacagcaca
     9061 agaactcaaa cggtagtgat atgaaaactg gtcccgcagc ccgttttata aagcccgctt
     9121 agcgctaaaa ccttatcagc cgcaatgctc tccgctctca agtcgcgttc aaaactttta
     9181 ccatatatgg gtatgaattg ttcgccgatt gggttttcat ttacggagaa tttcctccgc
     9241 aactgggtgt tcgtgcttgt gtgtgtcttg tatatcgcca tgaagagtgt acttccatgg
     9301 gtcctatcag ttgcggcatg tgattcaatt tgggggttac tcatattcga tacttcagct
     9361 ggttatcttt atgctgtgat ggtcacggtg cttttacaca ttttttcccc aactactcat
     9421 tgtatgccaa aacatacata tgtacatgtg tttatagtgg gtttattagt aatgtccagc
     9481 gatattaaaa cattacagtt aaataatatt catattagct tacgtttact gctatctgca
     9541 cttcaaattg gttattccag ggacattttg tgcaacacac atcttctagt agggcattca
     9601 ctagtcgtag aaaacacacc taacttaagc cggttttaac aataacggcg aaacagctgc
     9661 cagacgagag aaaagtacac agcaagaagc atttctataa tgatgccgtc ctaaatatgc
     9721 agcatttttg ctcaaattga tcatttcgtt gcatcgctca ggtggtatag tatttggcta
     9781 ttgaatgtga ttgctatctt aatagttttg aatagtaata gtgcaaatat tcattcataa
     9841 aaaataggta atcgattttt tcagtgcagt ctatgcgttt gcgcaacaga gacgaagaga
     9901 gagacgcctc acacacaagc acatgcggtc taagctgctc tccctctttt tctctctctc
     9961 tctggcttgc ggcagttttc cagaggctgt gactcagtaa gcatttcttg ttgttaccct
    10021 aacgataagc gtccgattcc taattctctt tcaaataatt tgttttattt ttttttttgt
    10081 gtgttttgaa gtcaaacagc ttcataaaca atattaaata tcgcaattgt tattgcaaat
    10141 gcattcgtat caggcgcctt gatgtttttc gccaaaactt cgtctgctct ttgtcagcgt
    10201 aattgcgttt gccggtcgtg ccgtcttgct cgcacgcggc ttgcaaacaa aaagtgaaag
    10261 agggaaagcc acctgtgttg ttgcttttgt tgctatcgtt ggctgcacac acatggcaat
    10321 aagtaaccat attgtagact tatttatttt agcgttttcg ttgactaatc tttgtttaaa
    10381 taattgccta tttcgagcca aaatgacgca taaaaggaaa cggcaacatg cggtgcgttg
    10441 agcatgcgta agcgagatgg cgctagagag agcgcagcat aaaaagaaaa aaacaaccgg
    10501 cgagtggcga gacacgaata atacacacac gcacacatgg tcaaaggagt ggctccaacg
    10561 tcggtgacgt cgcgcccagc cacagacccc gccacgcccc gccccaccct tccttccgct
    10621 gccccgcacc ccttt
//
