LOCUS       sequence-381227-        9512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  px458 with guide RNA that target hAAVS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9512)
  AUTHORS   Haiting Ma, Rudolf Jaenisch
  TITLE     Multiplex genome editing of human pluripotent stem cells using Cpf1
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 9512)
  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: "bioRxiv"
COMMENT     Sequence Label: pX458-AAVS1-sg
FEATURES             Location/Qualifiers
     source          1..9512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          228..455
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      467..476
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             476..541
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             548..568
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             593..4693
                     /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             593..4693
                     /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             4694..4741
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4757..4810
                     /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(4811..4813,4814..5524)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4811..4813,4814..5524)
                     /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(5114..5133)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5461..5482
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5552..5569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5558..5765
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5774..5914
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5774..5903
                     /label=AAV2 ITR
     rep_origin      5989..6444
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6076..6095)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6286..6307
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6461..6480)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6580..6602
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6640..6658)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6726..6830
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6831..6899,6900..7691)
                     /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(7049..7068)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7862..8450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8351..8370
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8512..8760
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8512..8532
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8683..8702
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8782..8857
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8954..9239
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cccccatatt ntdttttttt tttttttttk tattgttnng gggggggggg gggggggggg
       61 gggggggggg gcgcgcgcgn ggsggggcgg ggcggggcga ggggcggggc ggggcgaggc
      121 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
      181 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
      241 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
      301 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
      361 agctgagcaa gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt
      421 acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatggacta
      481 taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa
      541 gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa
      601 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
      661 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
      721 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
      781 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
      841 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
      901 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
      961 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     1021 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     1081 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     1141 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     1201 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     1261 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     1321 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     1381 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     1441 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     1501 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     1561 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     1621 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     1681 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     1741 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     1801 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     1861 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     1921 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     1981 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     2041 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     2101 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     2161 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     2221 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     2281 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     2341 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     2401 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     2461 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     2521 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     2581 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     2641 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     2701 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     2761 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     2821 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     2881 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     2941 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     3001 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     3061 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
     3121 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
     3181 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     3241 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
     3301 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     3361 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     3421 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     3481 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     3541 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     3601 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     3661 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     3721 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     3781 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     3841 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     3901 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     3961 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
     4021 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     4081 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     4141 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     4201 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     4261 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     4321 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     4381 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     4441 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     4501 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     4561 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
     4621 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     4681 gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa
     4741 ggaattcggc agtggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa
     4801 tcctggccca gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     4861 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     4921 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     4981 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     5041 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     5101 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     5161 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     5221 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     5281 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     5341 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     5401 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     5461 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     5521 caaggaattc taactagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     5581 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5641 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5701 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     5761 ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5821 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     5881 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     5941 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     6001 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6061 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6121 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6181 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     6241 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6301 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     6361 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6421 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     6481 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6541 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6601 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6661 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6721 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6781 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6841 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6901 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6961 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7021 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7081 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7141 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7201 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7261 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7321 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7381 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7441 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7501 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     7561 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7621 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7681 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7741 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7801 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7861 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7921 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7981 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     8041 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8101 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8161 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8221 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8281 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8341 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8401 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8461 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tgagggccta
     8521 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga
     8581 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
     8641 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
     8701 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
     8761 gggggccact agggacagga tgttttagag ctagaaatag caagttaaaa taaggctagt
     8821 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag agctagaaat
     8881 agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc agacaaatgg
     8941 ctctagaggt acccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     9001 acccccgccc attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg
     9061 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     9121 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc cagtacatga
     9181 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     9241 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
     9301 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
     9361 gsgcgcgccc rggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9421 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9481 gggggggggg gggggggggg gggggggggg gg
//
