LOCUS       sequence-262424-        9405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR based on px330 p2aGFP targeting TH gene near stop codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9405)
  AUTHORS   Ahfeldt T, Ordureau A, Bell C, Sarrafha L, Sun C, Piccinotti S, 
            Grass T, Parfitt GM, Paulo JA, Yanagawa F, Uozumi T, Kiyota Y, 
            Harper JW, Rubin LL
  TITLE     Pathogenic Pathways in Early-Onset Autosomal Recessive Parkinson's 
            Disease Discovered Using Isogenic Human Dopaminergic Neurons.
  JOURNAL   Stem Cell Reports. 2019 Dec 20. pii: S2213-6711(19)30444-8. doi: 
            10.1016/j.stemcr.2019.12.005.
  PUBMED    31902706
REFERENCE   2  (bases 1 to 9405)
  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: "Stem Cell 
            Reports. 2019 Dec 20. pii: S2213-6711(19)30444-8. doi: 
            10.1016/j.stemcr.2019.12.005."
COMMENT     Sequence Label: pTACR TH2-p2aGFP
FEATURES             Location/Qualifiers
     source          1..9405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        25..100
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        197..482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        484..761
                     /label=chicken beta-actin promoter
     intron          762..990
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1002..1011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1011..1076
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1083..1103
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1128..5228
                     /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             1128..5228
                     /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             5229..5276
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5277..5330
                     /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(5460..5462,5463..6176)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5460..5462,5463..6173)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5763..5782)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6110..6131
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6201..6218)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6207..6414
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6423..6563
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6423..6552
                     /label=AAV2 ITR
     rep_origin      6638..7093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6725..6744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6935..6956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7110..7129)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7229..7251
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7289..7307)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7375..7479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7480..7548,7549..8340)
                     /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(7698..7717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8511..9099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9000..9019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(9161..9405,1..4)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9161..9181
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9332..9351
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 cacctaggtg cacggcgtcc ctgagtttta gagctagaaa tagcaagtta aaataaggct
       61 agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga
      121 aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa
      181 tggctctaga ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      241 acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat
      301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca
      421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      481 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
      541 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
      601 gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
      661 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
      721 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgacgctgc
      781 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      841 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg
      901 agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg
      961 gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg gactataagg
     1021 accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg
     1081 ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca
     1141 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     1201 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     1261 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     1321 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     1381 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     1441 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     1501 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     1561 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     1621 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     1681 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     1741 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     1801 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     1861 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     1921 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     1981 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     2041 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     2101 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     2161 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     2221 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     2281 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     2341 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     2401 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     2461 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     2521 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     2581 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     2641 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     2701 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     2761 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     2821 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     2881 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     2941 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     3001 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     3061 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     3121 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     3181 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     3241 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     3301 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     3361 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     3421 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     3481 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     3541 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     3601 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     3661 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     3721 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     3781 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     3841 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     3901 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     3961 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     4021 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     4081 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     4141 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     4201 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     4261 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     4321 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     4381 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     4441 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     4501 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     4561 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     4621 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     4681 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     4741 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     4801 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     4861 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     4921 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     4981 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     5041 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     5101 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     5161 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     5221 gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggagg
     5281 gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca atgggttgct
     5341 gtttctccaa gaccgcagcg aagggagaag ccgccgcgga gaggcctggg gaggcggctg
     5401 tggcctcgtc gccttccaaa gcgaacggac aggagaatgg ccacgtgaag gtaggatccg
     5461 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     5521 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     5581 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     5641 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     5701 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     5761 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     5821 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     5881 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     5941 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     6001 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     6061 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     6121 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat
     6181 tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     6241 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     6301 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     6361 ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc
     6421 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     6481 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     6541 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     6601 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     6661 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6721 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6781 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6841 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6901 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6961 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     7021 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     7081 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     7141 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     7201 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     7261 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     7321 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     7381 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7441 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7501 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7561 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7621 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7681 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7741 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7801 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7861 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7921 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7981 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     8041 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     8101 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     8161 tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg
     8221 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     8281 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     8341 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     8401 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     8461 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     8521 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     8581 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     8641 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     8701 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8761 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8821 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8881 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8941 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     9001 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     9061 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     9121 ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat ttcccatgat
     9181 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
     9241 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     9301 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     9361 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaa
//
