LOCUS       sequence-267550-        9401 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 9401)
  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 9401)
  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..9401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        45..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     gap             444..453
                     /estimated_length=10
     intron          607..834
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      846..855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             855..920
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             927..947
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             972..5072
                     /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             972..5072
                     /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             5073..5120
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5121..5174
                     /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(5304..5306,5307..6020)
                     /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(5304..5306,5307..6017)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5607..5626)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5954..5975
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6045..6062)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6051..6258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6267..6407
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6267..6396
                     /label=AAV2 ITR
     rep_origin      6482..6937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6569..6588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6779..6800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6954..6973)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7073..7095
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7133..7151)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7219..7323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7324..7392,7393..8184)
                     /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(7542..7561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8355..8943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8844..8863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        9005..9253
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9005..9025
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9176..9195
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9274..9349
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tagcgcgtgc gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg
       61 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
      361 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
      421 tgtgcagcga tgggggcggg gggnnnnnnn nnngcgccag gcggggcggg gcggggcgag
      481 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
      541 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      601 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      661 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      721 tctcctccgg gctgtaatta gctgagcaag aggtaagggt ttaagggatg gttggttggt
      781 ggggtattaa tgtttaatta cctggagcac ctgcctgaaa tcactttttt tcaggttgga
      841 ccggtgccac catggactat aaggaccacg acggagacta caaggatcat gatattgatt
      901 acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag
      961 tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct
     1021 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     1081 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     1141 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     1201 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     1261 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     1321 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     1381 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     1441 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     1501 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     1561 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     1621 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     1681 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     1741 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     1801 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     1861 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     1921 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     1981 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     2041 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     2101 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     2161 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     2221 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     2281 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     2341 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     2401 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     2461 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     2521 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     2581 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     2641 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     2701 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     2761 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     2821 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     2881 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     2941 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     3001 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     3061 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     3121 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     3181 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     3241 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     3301 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     3361 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     3421 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     3481 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     3541 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     3601 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     3661 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     3721 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     3781 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     3841 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     3901 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     3961 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     4021 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     4081 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     4141 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     4201 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     4261 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     4321 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     4381 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     4441 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     4501 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     4561 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     4621 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     4681 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     4741 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     4801 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     4861 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     4921 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     4981 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     5041 cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg
     5101 gccaggcaaa aaagaaaaag gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     5161 agaatcctgg cccaatgggt tgctgtttct ccaagaccgc agcgaaggga gaagccgccg
     5221 cggagaggcc tggggaggcg gctgtggcct cgtcgccttc caaagcgaac ggacaggaga
     5281 atggccacgt gaaggtagga tccgtgagca agggcgagga gctgttcacc ggggtggtgc
     5341 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     5401 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     5461 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     5521 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     5581 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     5641 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     5701 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     5761 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     5821 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     5881 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     5941 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     6001 tggacgagct gtacaagtaa gaattcctag agctcgctga tcagcctcga ctgtgccttc
     6061 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     6121 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     6181 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa
     6241 tagcaggcat gctggggagc ggccgcagga acccctagtg atggagttgg ccactccctc
     6301 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     6361 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     6421 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     6481 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6541 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6601 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6661 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     6721 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6781 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     6841 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     6901 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     6961 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     7021 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     7081 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     7141 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     7201 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7261 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7321 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7381 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7441 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7501 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7561 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7621 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7681 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7741 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7801 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7861 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7921 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7981 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga
     8041 agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8101 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8161 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8221 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8281 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8341 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8401 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8461 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8521 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8581 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8641 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8701 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8761 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8821 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8881 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8941 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     9001 atgtgagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     9061 agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     9121 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     9181 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     9241 aggacgaaac acctaggtgc acggcgtccc tgagttttag agctagaaat agcaagttaa
     9301 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttt
     9361 agagctagaa atagcaagtt aaaataaggc tagtccgttt t
//
