LOCUS       sequence-267552-        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 TH1-p2aGFP
FEATURES             Location/Qualifiers
     source          1..9401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          158..385
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      397..406
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             406..471
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             478..498
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             523..4623
                     /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             523..4623
                     /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             4624..4671
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4672..4725
                     /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(4855..4857,4858..5571)
                     /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(4855..4857,4858..5568)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5158..5177)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5505..5526
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5596..5613)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5602..5809
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5818..5958
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5818..5947
                     /label=AAV2 ITR
     rep_origin      6033..6488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6120..6139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6330..6351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6505..6524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6624..6646
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6684..6702)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6770..6874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6875..6943,6944..7735)
                     /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(7093..7112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7906..8494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8395..8414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8556..8804
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8556..8576
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8727..8746
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8825..8900
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8997..9282
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(9284..9401,1..157)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc
       61 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
      121 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg
      181 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
      241 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa
      301 gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca
      361 cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatggacta taaggaccac
      421 gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca
      481 aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc
      541 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
      601 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
      661 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
      721 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
      781 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
      841 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
      901 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
      961 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     1021 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     1081 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     1141 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     1201 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     1261 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     1321 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     1381 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     1441 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     1501 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     1561 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     1621 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     1681 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     1741 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     1801 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     1861 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     1921 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     1981 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     2041 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     2101 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     2161 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     2221 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     2281 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     2341 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     2401 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     2461 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     2521 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     2581 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     2641 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     2701 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     2761 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     2821 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     2881 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     2941 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     3001 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
     3061 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
     3121 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
     3181 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
     3241 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     3301 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
     3361 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     3421 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     3481 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     3541 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     3601 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     3661 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     3721 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
     3781 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     3841 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
     3901 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     3961 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     4021 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     4081 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     4141 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     4201 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     4261 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
     4321 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     4381 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
     4441 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     4501 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
     4561 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
     4621 gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa ggagggcaga
     4681 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaatggg ttgctgtttc
     4741 tccaagaccg cagcgaaggg agaagccgcc gcggagaggc ctggggaggc ggctgtggcc
     4801 tcgtcgcctt ccaaagcgaa cggacaggag aatggccacg tgaaggtagg atccgtgagc
     4861 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     4921 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     4981 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     5041 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     5101 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     5161 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     5221 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     5281 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     5341 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     5401 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     5461 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     5521 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agaattccta
     5581 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5641 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5701 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5761 aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag cggccgcagg
     5821 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     5881 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     5941 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6001 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     6061 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     6121 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     6181 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     6241 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     6301 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     6361 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     6421 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     6481 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     6541 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     6601 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     6661 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     6721 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6781 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6841 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6901 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6961 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7021 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7081 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7141 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     7201 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     7261 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     7321 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     7381 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     7441 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     7501 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     7561 tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag cactggggcc
     7621 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7681 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7741 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7801 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7861 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7921 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7981 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8041 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     8101 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     8161 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     8221 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     8281 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     8341 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8401 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8461 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8521 gttcctggcc ttttgctggc cttttgctca catgtgaggg cctatttccc atgattcctt
     8581 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
     8641 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
     8701 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
     8761 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgacgcc gtgcacctag
     8821 ccaagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
     8881 aagtggcacc gagtcggtgc ttttttgttt tagagctaga aatagcaagt taaaataagg
     8941 ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa tggctctaga ggtacccgtt
     9001 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     9061 tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     9121 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     9181 aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct
     9241 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     9301 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     9361 ttttaattat tttgtgcagc gatgggggcg gggggggggg g
//
