LOCUS       sequence-373464-       10635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses scrambled sgRNA 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.scr_1
FEATURES             Location/Qualifiers
     source          1..10635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        835..1234
                     /label=dU6-2 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-2 
                     snRNA (Das et al., 1987)"
     misc_RNA        1256..1331
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1483..1501)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1569..1673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1674..1742,1743..2534)
                     /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(1892..1911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2705..3293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3194..3213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3894..4028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3931..3950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3985..4004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4036..4635)
                     /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(4119..4139)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4616..4635
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4660..4713)
                     /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(4729..4776)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4777..8877)
                     /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(4777..8877)
                     /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(8902..8922)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8929..8994)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 gccacgcccc gccccaccct tccttccgct gccccgcacc cctttccttc cactaccgtt
       61 ttgccgcatt tggtattgcc atctcgcttt tttggcttta ctttgctttt tgaataagcc
      121 aagaaactga aatcaaagcg gacagtccga aaattgaata ttccaaaaaa caacaattgc
      181 acatgcactg cgttcggcaa aagacgcaga aattcaagcc ttttggtctg gcatcatctt
      241 aaaatatcat cagtatcatc attgccatta taaatacata agaaaatgag aatacattaa
      301 agatatacat acaatctttg tcatagtcaa catattttgc aacaaaatta tctatagata
      361 acatgttttt gtagaaaatt ccactaactg attttcgtat aaatacaaat tattctcatt
      421 cttcaaaata ctcgccgacg tcatcgcgaa attcctcagc attcttcgac aaatgaagag
      481 agacagcgag tgacacaatc ttatcttgcc gacgagtctt tgaaaatggc cccgggtaac
      541 ggtgaaaaac atatagccaa gaatggagcg atcgccaggc gatggaaatt ggcttcaaat
      601 tgctccgaat atggccagtt ggtcagttgg cattggattg tggagagata ctagatcagc
      661 tcgttctcct tctcctccgc attctcctac tccgcccgaa gacgcttctt tgatcgcttc
      721 tttctcgaat gtgtacatat tcattatgta tttggtgttt ggcgacgatc agccagcttt
      781 aatggccata aaacgtgtct tggtttgtgg gccacataac aggcagcgag atctgttcga
      841 cttgcagcct gaaatacggc acgagtagga aaagccgagt caaatgccga atgcagagtc
      901 tcattacagc acaatcaact caagaaaaac tcgacacttt tttaccattt gcacttaaat
      961 ccttttttat tcgttatgta tacttttttt ggtccctaac caaaacaaaa ccaaactctc
     1021 ttagtcgtgc ctctatattt aaaactatca atttattata gtcaataaat cgaactgtgt
     1081 tttcaacaaa cgaacaatag gacactttga ttctaaagga aattttgaaa atcttaagca
     1141 gagggttctt aagaccattt gccaattctt ataattctca actgctcttt cctgatgttg
     1201 atcatttata taggtatgtt ttcctcaata cttcgctggg ctactccagt aaggagtttt
     1261 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     1321 cgagtcggtg cttttttaga tctggaaaaa tgatgtgaca gtggaaatga gaagtaggtg
     1381 catctgcaaa gatttgattc agagttgatg ccattcatga ttttttttta gaagtatcat
     1441 ttgtgttttt agaatataga attaattctt gaagacgaaa gggcctcgtg atacgcctat
     1501 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1561 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1621 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1681 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1741 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1801 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1861 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1921 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1981 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2041 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2101 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2161 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     2221 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2281 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2341 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2401 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2461 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2521 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2581 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2641 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2701 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2761 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2821 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2881 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2941 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3001 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3061 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3121 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3181 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3241 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3301 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttgaagctgt ccctgatggt
     3361 cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag
     3421 cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt
     3481 agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     3541 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     3601 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     3661 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     3721 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     3781 gtcgactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3841 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3901 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3961 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4021 tctggatccc tcgagtcagg caccgggctt gcgggtcatg caccaggtgc gcggtccttc
     4081 gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg
     4141 gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg
     4201 gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc
     4261 caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg
     4321 cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc
     4381 ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac
     4441 ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac
     4501 ccgctcgatg tggcggtccg ggtcgacggt gtggcgcgtg gcggggtagt cggcgaacgc
     4561 ggcggcgagg gtgcgtacgg cccgggggac gtcgtcgcgg gtggcgaggc gcaccgtggg
     4621 cttgtactcg gtcatgctac cgctgccgct accgctagca gggccgggat tctcctccac
     4681 gtcaccgcat gttagaagac ttcctctgcc ctcaagcttg cctgatccct ttttcttttt
     4741 tgcctggccg gcctttttcg tggccgccgg ccttttgtcg cctcccagct gagacaggtc
     4801 gatccgtgtc tcgtacaggc cggtgatgct ctggtggatc agggtggcgt ccagcacctc
     4861 tttggtgctg gtgtacctct tccggtcgat ggtggtgtca aagtacttga aggcggcagg
     4921 ggctcccaga ttggtcaggg taaacaggtg gatgatattc tcggcctgct ctctgatggg
     4981 cttatcccgg tgcttgttgt aggcggacag cactttgtcc agattagcgt cggccaggat
     5041 cactctcttg gagaactcgc tgatctgctc gatgatctcg tccaggtagt gcttgtgctg
     5101 ttccacaaac agctgtttct gctcattatc ctcgggggag cccttcagct tctcatagtg
     5161 gctggccagg tacaggaagt tcacatattt ggagggcagg gccagttcgt ttcccttctg
     5221 cagttcgccg gcagaggcca gcattctctt ccggccgttt tccagctcga acagggagta
     5281 cttaggcagc ttgatgatca ggtccttttt cacttctttg tagcccttgg cttccagaaa
     5341 gtcgatggga ttcttctcga agctgcttct ttccatgatg gtgatcccca gcagctcttt
     5401 cacactcttc agtttcttgg acttgccctt ttccactttg gccaccacca gcacagaata
     5461 ggccacggtg gggctgtcga agccgccgta cttcttaggg tcccagtcct tctttctggc
     5521 gatcagctta tcgctgttcc tcttgggcag gatagactct ttgctgaagc cgcctgtctg
     5581 cacctcggtc tttttcacga tattcacttg gggcatgctc agcactttcc gcacggtggc
     5641 aaaatcccgg cccttatccc acacgatctc cccggtttcg ccgtttgtct cgatcagagg
     5701 ccgcttccgg atctcgccgt tggccagggt aatctcggtc ttgaaaaagt tcatgatgtt
     5761 gctgtagaag aagtacttgg cggtagcctt gccgatttcc tgctcgctct tggcgatcat
     5821 cttccgcacg tcgtacacct tgtagtcgcc gtacacgaac tcgctttcca gcttagggta
     5881 ctttttgatc agggcggttc ccacgacggc gttcaggtag gcgtcgtggg cgtggtggta
     5941 gttgttgatc tcgcgcactt tgtaaaactg gaaatccttc cggaaatcgg acaccagctt
     6001 ggacttcagg gtgatcactt tcacttcccg gatcagcttg tcattctcgt cgtacttagt
     6061 gttcatccgg gagtccagga tctgtgccac gtgctttgtg atctgccggg tttccaccag
     6121 ctgtctcttg atgaagccgg ccttatccag ttcgctcagg ccgcctctct cggccttggt
     6181 cagattgtcg aactttctct gggtaatcag cttggcgttc agcagctgcc gccagtagtt
     6241 cttcatcttc ttcacgacct cttcggaggg cacgttgtcg ctcttgcccc ggttcttgtc
     6301 gcttctggtc agcaccttgt tgtcgatgga gtcgtccttc agaaagctct gaggcacgat
     6361 atggtccaca tcgtagtcgg acagccggtt gatgtccagt tcctggtcca cgtacatatc
     6421 ccgcccattc tgcaggtagt acaggtacag cttctcgttc tgcagctggg tgttttccac
     6481 ggggtgttct ttcaggatct ggctgcccag ctctttgatg ccctcttcga tccgcttcat
     6541 tctctcgcgg ctgttcttct gtcccttctg ggtggtctgg ttctctctgg ccatttcgat
     6601 cacgatgttc tcgggcttgt gccggcccat cactttcacg agctcgtcca ccaccttcac
     6661 tgtctgcagg atgcccttct taatggcggg gctgccggcc agattggcaa tgtgctcgtg
     6721 caggctatcg ccctggccgg acacctgggc tttctggatg tcctctttaa aggtcaggct
     6781 gtcgtcgtgg atcagctgca tgaagtttct gttggcgaag ccgtcggact tcaggaaatc
     6841 caggattgtc ttgccggact gcttgtcccg gatgccgttg atcagcttcc ggctcagcct
     6901 gccccagccg gtgtatctcc gccgcttcag ctgcttcatc actttgtcgt cgaacaggtg
     6961 ggcataggtt ttcagccgtt cctcgatcat ctctctgtcc tcaaacagtg tcagggtcag
     7021 cacgatatct tccagaatgt cctcgttttc ctcattgtcc aggaagtcct tgtccttgat
     7081 aattttcagc agatcgtggt atgtgcccag ggaggcgttg aaccgatctt ccacgccgga
     7141 gatttccacg gagtcgaagc actcgatttt cttgaagtag tcctctttca gctgcttcac
     7201 ggtcactttc cggttggtct tgaacagcag gtccacgatg gcctttttct gctcgccgct
     7261 caggaaggcg ggctttctca ttccctcggt cacgtatttc actttggtca gctcgttata
     7321 cacggtgaag tactcgtaca gcaggctgtg cttgggcagc accttctcgt tgggcaggtt
     7381 cttatcgaag ttggtcatcc gctcgatgaa gctctgggcg gaagcgccct tgtccaccac
     7441 ttcctcgaag ttccaggggg tgatggtttc ctcgctcttt ctggtcatcc aggcgaatct
     7501 gctgtttccc ctggccagag ggcccacgta gtaggggatg cggaaggtca ggatcttctc
     7561 gatcttttcc cggttgtcct tcaggaatgg gtaaaaatct tcctgccgcc gcagaatggc
     7621 gtgcagctct cccaggtgga tctggtgggg gatgctgccg ttgtcgaagg tccgctgctt
     7681 ccgcagcagg tcctctctgt tcagcttcac gagcagttcc tcggtgccgt ccatcttttc
     7741 caggatgggc ttgatgaact tgtagaactc ttcctggctg gctccgccgt caatgtagcc
     7801 ggcgtagccg ttcttgctct ggtcgaagaa aatctctttg tacttctcag gcagctgctg
     7861 ccgcacgaga gctttcagca gggtcaggtc ctggtggtgc tcgtcgtatc tcttgatcat
     7921 agaggcgctc aggggggcct tggtgatctc ggtgttcact ctcaggatgt cgctcagcag
     7981 gatggcgtcg gacaggttct tggcggccag aaacaggtcg gcgtactggt cgccgatctg
     8041 ggccagcagg ttgtccaggt cgtcgtcgta ggtgtccttg ctcagctgca gtttggcatc
     8101 ctcggccagg tcgaagttgc tcttgaagtt gggggtcagg cccaggctca gggcaatcag
     8161 gtttccgaac aggccattct tcttctcgcc gggcagctgg gcgatcagat tttccagccg
     8221 tctgctcttg ctcagtctgg cagacaggat ggccttggcg tccacgccgc tggcgttgat
     8281 ggggttttcc tcgaacagct ggttgtaggt ctgcaccagc tggatgaaca gcttgtccac
     8341 gtcgctgttg tcggggttca ggtcgccctc gatcaggaag tggccccgga acttgatcat
     8401 gtgggccagg gccagataga tcagccgcag gtcggccttg tcggtgctgt ccaccagttt
     8461 ctttctcagg tggtagatgg tggggtactt ctcgtggtag gccacctcgt ccacgatgtt
     8521 gccgaagatg gggtgccgct cgtgcttctt atcctcttcc accaggaagg actcttccag
     8581 tctgtggaag aagctgtcgt ccaccttggc catctcgttg ctgaagatct cttgcagata
     8641 gcagatccgg ttcttccgtc tggtgtatct tcttctggcg gttctcttca gccgggtggc
     8701 ctcggctgtt tcgccgctgt cgaacagcag ggctccgatc aggttcttct tgatgctgtg
     8761 ccggtcggtg ttgcccagca ccttgaattt cttgctgggc accttgtact cgtcggtgat
     8821 cacggcccag cccacagagt tggtgccgat gtccaggccg atgctgtact tcttgtcggc
     8881 tgctgggact ccgtggatac cgaccttccg cttcttcttt ggggccatct tatcgtcatc
     8941 gtctttgtaa tcaatatcat gatccttgta gtctccgtcg tggtccttat agtccatgaa
     9001 ttccaccaca ctggactagt aggtaccccg atccggggtc tctggattag acgactgctg
     9061 gctgatggag cggctttgtg tcgggaggag tatccacaca gcacaagaac tcaaacggta
     9121 gtgatatgaa aactggtccc gcagcccgtt ttataaagcc cgcttagcgc taaaacctta
     9181 tcagccgcaa tgctctccgc tctcaagtcg cgttcaaaac ttttaccata tatgggtatg
     9241 aattgttcgc cgattgggtt ttcatttacg gagaatttcc tccgcaactg ggtgttcgtg
     9301 cttgtgtgtg tcttgtatat cgccatgaag agtgtacttc catgggtcct atcagttgcg
     9361 gcatgtgatt caatttgggg gttactcata ttcgatactt cagctggtta tctttatgct
     9421 gtgatggtca cggtgctttt acacattttt tccccaacta ctcattgtat gccaaaacat
     9481 acatatgtac atgtgtttat agtgggttta ttagtaatgt ccagcgatat taaaacatta
     9541 cagttaaata atattcatat tagcttacgt ttactgctat ctgcacttca aattggttat
     9601 tccagggaca ttttgtgcaa cacacatctt ctagtagggc attcactagt cgtagaaaac
     9661 acacctaact taagccggtt ttaacaataa cggcgaaaca gctgccagac gagagaaaag
     9721 tacacagcaa gaagcatttc tataatgatg ccgtcctaaa tatgcagcat ttttgctcaa
     9781 attgatcatt tcgttgcatc gctcaggtgg tatagtattt ggctattgaa tgtgattgct
     9841 atcttaatag ttttgaatag taatagtgca aatattcatt cataaaaaat aggtaatcga
     9901 ttttttcagt gcagtctatg cgtttgcgca acagagacga agagagagac gcctcacaca
     9961 caagcacatg cggtctaagc tgctctccct ctttttctct ctctctctgg cttgcggcag
    10021 ttttccagag gctgtgactc agtaagcatt tcttgttgtt accctaacga taagcgtccg
    10081 attcctaatt ctctttcaaa taatttgttt tatttttttt tttgtgtgtt ttgaagtcaa
    10141 acagcttcat aaacaatatt aaatatcgca attgttattg caaatgcatt cgtatcaggc
    10201 gccttgatgt ttttcgccaa aacttcgtct gctctttgtc agcgtaattg cgtttgccgg
    10261 tcgtgccgtc ttgctcgcac gcggcttgca aacaaaaagt gaaagaggga aagccacctg
    10321 tgttgttgct tttgttgcta tcgttggctg cacacacatg gcaataagta accatattgt
    10381 agacttattt attttagcgt tttcgttgac taatctttgt ttaaataatt gcctatttcg
    10441 agccaaaatg acgcataaaa ggaaacggca acatgcggtg cgttgagcat gcgtaagcga
    10501 gatggcgcta gagagagcgc agcataaaaa gaaaaaaaca accggcgagt ggcgagacac
    10561 gaataataca cacacgcaca catggtcaaa ggagtggctc caacgtcggt gacgtcgcgc
    10621 ccagccacag acccc
//
