LOCUS       sequence-328128-        9826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 expressing plasmid containing the gRNA targeting an 
            enhancer of mouse Otx2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9826)
  AUTHORS   Kaufman ML, Goodson NB, Park KU, Schwanke M, Office E, Schneider SR,
            Abraham J, Hensley A, Jones KL, Brzezinski JA
  TITLE     Initiation of Otx2 expression in the developing mouse retina 
            requires a unique enhancer and either Ascl1 or Neurog2 activity.
  JOURNAL   Development. 2021 Jun 15;148(12). pii: 269198. doi: 
            10.1242/dev.199399. Epub 2021 Jun 18.
  PUBMED    34143204
REFERENCE   2  (bases 1 to 9826)
  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: 
            "Development."; date: "2021-06-15"; volume: "148(12). pii: 269198. 
            doi"; pages: " 10.1242/dev.199399. Epub 2021 Jun 18"
COMMENT     Sequence Label: pX458-Ef1a-Cas9-GFP (gRNA: DHS-4D-1)
FEATURES             Location/Qualifiers
     source          1..9826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        90..1272
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          321..1263
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1220..1240
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1278..1287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1287..1352
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1359..1379
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1404..5504
                     /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             1404..5504
                     /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             5505..5552
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5568..5621
                     /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             join(5622..5624,5625..6335)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5622..5624,5625..6335)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5925..5944)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6272..6293
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6363..6380)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6369..6576
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6585..6725
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6585..6714
                     /label=AAV2 ITR
     rep_origin      6800..7255
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6887..6906)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7097..7118
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7272..7291)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7391..7413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7451..7469)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7537..7641
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7642..7710,7711..8502)
                     /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(7860..7879)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8673..9261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9162..9181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        9323..9571
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9323..9343
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9494..9513
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9593..9668
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ccactaacgt agaacccaga gatcgctgcg ttcccgcccc ctcacccgcc cgctctcgtc
       61 atcactgagg tggagaagag catgcgtgag gctccggtgc ccgtcagtgg gcagagcgca
      121 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
      181 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      241 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      301 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
      361 cgggttatgg cccttgcgtg ccttgaatta cttccacgcc cctggctgca gtacgtgatt
      421 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
      481 gccccttcgc ctcgtgcttg agttgaggcc tggcttgggc gctggggccg ccgcgtgcga
      541 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
      601 ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
      661 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
      721 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
      781 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
      841 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
      901 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
      961 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     1021 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     1081 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     1141 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     1201 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     1261 caggtgtcgt gaccggtgcc accatggact ataaggacca cgacggagac tacaaggatc
     1321 atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg
     1381 gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca
     1441 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
     1501 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg
     1561 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     1621 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     1681 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     1741 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     1801 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     1861 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     1921 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     1981 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     2041 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     2101 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     2161 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     2221 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     2281 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     2341 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     2401 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     2461 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     2521 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     2581 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     2641 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     2701 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     2761 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     2821 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
     2881 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     2941 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
     3001 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
     3061 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     3121 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     3181 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     3241 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     3301 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     3361 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     3421 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     3481 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     3541 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     3601 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     3661 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     3721 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     3781 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
     3841 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
     3901 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca
     3961 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
     4021 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
     4081 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
     4141 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
     4201 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
     4261 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
     4321 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
     4381 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
     4441 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
     4501 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
     4561 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
     4621 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
     4681 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
     4741 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
     4801 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
     4861 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
     4921 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
     4981 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
     5041 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
     5101 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
     5161 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
     5221 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
     5281 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
     5341 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
     5401 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
     5461 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca
     5521 cgaaaaaggc cggccaggca aaaaagaaaa aggaattcgg cagtggagag ggcagaggaa
     5581 gtctgctaac atgcggtgac gtcgaggaga atcctggccc agtgagcaag ggcgaggagc
     5641 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     5701 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     5761 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     5821 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     5881 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     5941 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     6001 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     6061 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     6121 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     6181 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     6241 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     6301 ccgggatcac tctcggcatg gacgagctgt acaaggaatt ctaactagag ctcgctgatc
     6361 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     6421 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6481 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     6541 ggaggattgg gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat
     6601 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     6661 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     6721 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     6781 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6841 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     6901 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6961 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     7021 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     7081 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     7141 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     7201 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     7261 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     7321 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     7381 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     7441 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     7501 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     7561 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     7621 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     7681 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     7741 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     7801 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     7861 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     7921 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7981 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     8041 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     8101 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     8161 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     8221 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     8281 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     8341 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     8401 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     8461 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     8521 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     8581 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8641 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     8701 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     8761 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     8821 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     8881 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     8941 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     9001 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     9061 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     9121 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     9181 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     9241 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     9301 tgctggcctt ttgctcacat gtgagggcct atttcccatg attccttcat atttgcatat
     9361 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt
     9421 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
     9481 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
     9541 tttatatatc ttgtggaaag gacgaaacac cgattacagt aggagcgagg gggttttaga
     9601 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     9661 gtcggtgctt ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt
     9721 tagcgcgtgc gccaattctg cagacaaatg gctctagagg taccagtaat tcatacaaaa
     9781 ggactcgccc ctgccttggg gaatcccagg gaccgtcgtt aaactc
//
