LOCUS       sequence-405307-        9276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid of dead SpCas9 with T2A-EGFP and 
            cloning backbone for sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9276)
  AUTHORS   Acharya S, Ansari AH, Kumar Das P, Hirano S, Aich M, Rauthan R, 
            Mahato S, Maddileti S, Sarkar S, Kumar M, Phutela R, Gulati S, 
            Rahman A, Goel A, Afzal C, Paul D, Agrawal T, Pulimamidi VK, Jalali 
            S, Nishimasu H, Mariappan I, Nureki O, Maiti S, Chakraborty D
  TITLE     PAM-flexible Engineered FnCas9 variants for robust and ultra-precise
            genome editing and diagnostics.
  JOURNAL   Nat Commun. 2024 Jun 28;15(1):5471. doi: 10.1038/s41467-024-49233-w.
  PUBMED    38942756
REFERENCE   2  (bases 1 to 9276)
  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: "Nat 
            Commun."; date: "2024-06-28"; volume: "15(1):5471. doi"; pages: " 
            10.1038/s41467-024-49233-w"
COMMENT     Sequence Label: PX458-3xHA-dSpCas9
FEATURES             Location/Qualifiers
     source          1..9276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        268..343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        440..725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          972..1199
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1211..1220
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1226..1306
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             1313..1333
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1358..5458
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPEYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5459..5506
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5522..5575
                     /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(5576..5578,5579..6289)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5576..5578,5579..6289)
                     /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(5879..5898)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6226..6247
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6317..6334)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6323..6530
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6539..6679
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6539..6668
                     /label=AAV2 ITR
     rep_origin      6754..7209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6841..6860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7051..7072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7226..7245)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7345..7367
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7405..7423)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7491..7595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7596..7664,7665..8456)
                     /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(7814..7833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8627..9215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9116..9135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
      541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
      661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcggggcggg
      841 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
      901 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
      961 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
     1021 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
     1081 gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg
     1141 ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg
     1201 ttggaccggt gccaccatgg gatcctaccc atacgatgtt ccagattacg cttacccata
     1261 cgatgttcca gattacgctt acccatacga tgttccagat tacgctatgg ccccaaagaa
     1321 gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct
     1381 ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     1441 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     1501 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     1561 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     1621 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     1681 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     1741 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     1801 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     1861 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     1921 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     1981 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     2041 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     2101 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     2161 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     2221 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     2281 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     2341 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     2401 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     2461 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     2521 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     2581 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     2641 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     2701 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     2761 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     2821 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     2881 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     2941 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     3001 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     3061 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     3121 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     3181 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     3241 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     3301 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     3361 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     3421 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     3481 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     3541 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     3601 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     3661 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     3721 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     3781 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     3841 catcaaccgg ctgtccgact acgatgtgga cgccatcgtg cctcagagct ttctgaagga
     3901 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     3961 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     4021 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     4081 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     4141 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     4201 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     4261 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     4321 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     4381 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     4441 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     4501 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     4561 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     4621 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     4681 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     4741 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     4801 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     4861 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     4921 agtgaaaaag gacctgatca tcaagctgcc tgagtactcc ctgttcgagc tggaaaacgg
     4981 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     5041 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     5101 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     5161 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     5221 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     5281 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     5341 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     5401 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa
     5461 aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg
     5521 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag
     5581 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     5641 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     5701 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5761 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     5821 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     5881 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5941 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     6001 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     6061 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     6121 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     6181 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     6241 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaact
     6301 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6361 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     6421 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     6481 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag
     6541 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     6601 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     6661 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     6721 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     6781 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     6841 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6901 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6961 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     7021 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     7081 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     7141 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     7201 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     7261 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     7321 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     7381 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     7441 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     7501 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     7561 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     7621 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     7681 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     7741 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7801 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7861 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7921 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7981 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     8041 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     8101 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     8161 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     8221 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     8281 ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc
     8341 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     8401 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     8461 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     8521 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     8581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     8641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     8701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8761 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     9001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     9061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     9121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     9181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     9241 ggttcctggc cttttgctgg ccttttgctc acatgt
//
