LOCUS       sequence-127248-       10323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human codon optimized Csy4 and FokI-dCas9 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10323)
  AUTHORS   Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin
            MJ, Aryee MJ, Joung JK
  TITLE     Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome 
            editing.
  JOURNAL   Nat Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2908.
  PUBMED    24770325
REFERENCE   2  (bases 1 to 10323)
  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 
            Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2908."
COMMENT     Sequence Label: pSQT1601
FEATURES             Location/Qualifiers
     source          1..10323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1989..2368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          2622..3638
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3562..3584
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3646..3665
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3731..3740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4301..4354
                     /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             4358..4378
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4385..4972
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     CDS             join(4988..8386,8387..8389,8390..8986,8987,8988..8990,
                     8991..8992,8993..8995,8996..9088)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(4988..5011,5012..5014,5015..7501,7502..7504,
                     7505..9088)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9095..9115
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9122..9187
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(9277..9296)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9342..9397
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9396..9415)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(9758..9774)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9758..9774)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9771..9793)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9782..9798
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9806..9812,9813..9830,9831..9836))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9851..9872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9931..10126
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9977..10112
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    10132..10266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10169..10188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10223..10242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     2041 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     2101 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     2161 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     2221 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     2281 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     2341 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     2401 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     2461 tgcagcgatg ggggcggggg gggggggggg gcgaggggcg gggcggggcg aggcggagag
     2521 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
     2581 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
     2641 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
     2701 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
     2761 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
     2821 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
     2881 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
     2941 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
     3001 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     3061 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
     3121 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
     3181 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     3241 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
     3301 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     3361 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     3421 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     3481 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     3541 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     3601 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     3661 ttgtgctgtc tcatcatttt ggcaaagaat tctgcagtcg acggtaccgc gggcccggga
     3721 tccaccggtc gccaccatgg gtgatcatta tctggatatt cggctgaggc ctgatccaga
     3781 gttcccacct gcgcagctga tgtctgtcct ttttggcaaa cttcatcagg ccctggttgc
     3841 ccagggcgga gatcggatag gggtaagctt tccagacctc gacgaaagcc ggagccgcct
     3901 gggagaacgc ctgcggatcc acgcttctgc cgacgatctg agagccttgc tggcaaggcc
     3961 atggcttgag gggctccggg atcacctgca gtttggcgaa cccgccgttg ttccccaccc
     4021 aaccccttat cggcaggtgt ctagagtgca ggccaaatct aatccagaac ggctgcgacg
     4081 gcgactcatg cggcgacatg atcttagcga ggaagaggcc cgaaaaagaa tccctgatac
     4141 cgtggcccgc gcccttgact tgccttttgt cacactgcgg tcccagagta cggggcagca
     4201 tttcagactt ttcattcgac acgggccact gcaagttacc gccgaagaag gaggctttac
     4261 ttgttatgga ctctccaagg gaggtttcgt gccctggttt gagggcagag gaagtctgtt
     4321 aacatgcggt gacgtcgagg agaatcctgg cccaatgcct aagaagaagc ggaaggtgag
     4381 cagccaactt gtgaagtctg aactcgagga gaaaaaatca gagttgagac acaagttgaa
     4441 gtacgtgcca cacgaataca tcgagcttat cgagatcgcc agaaacagta cccaggatag
     4501 gatccttgag atgaaagtca tggagttctt tatgaaggtc tacggttata gaggaaagca
     4561 ccttggcggt agcagaaagc ccgatggcgc catctatact gtcggatctc ctatcgatta
     4621 tggggtgatc gtggatacca aagcttactc aggcgggtac aacttgccca taggacaagc
     4681 cgacgagatg cagcggtatg tcgaagagaa ccagacgcgc aacaagcaca tcaaccccaa
     4741 tgaatggtgg aaagtgtacc caagtagtgt gactgagttc aagttcctgt ttgtctccgg
     4801 ccactttaag ggcaattata aagctcagct cactagactc aatcacatca caaactgcaa
     4861 cggagctgtg ttgtcagtgg aggagctcct gattggaggc gagatgatca aagccggcac
     4921 ccttacactg gaggaggtgc ggcggaagtt caacaatgga gagatcaact tcggtggcgg
     4981 tggatccgat aaaaagtatt ctattggttt agccatcggc actaattccg ttggatgggc
     5041 tgtcataacc gatgaataca aagtaccttc aaagaaattt aaggtgttgg ggaacacaga
     5101 ccgtcattcg attaaaaaga atcttatcgg tgccctccta ttcgatagtg gcgaaacggc
     5161 agaggcgact cgcctgaaac gaaccgctcg gagaaggtat acacgtcgca agaaccgaat
     5221 atgttactta caagaaattt ttagcaatga gatggccaaa gttgacgatt ctttctttca
     5281 ccgtttggaa gagtccttcc ttgtcgaaga ggacaagaaa catgaacggc accccatctt
     5341 tggaaacata gtagatgagg tggcatatca tgaaaagtac ccaacgattt atcacctcag
     5401 aaaaaagcta gttgactcaa ctgataaagc ggacctgagg ttaatctact tggctcttgc
     5461 ccatatgata aagttccgtg ggcactttct cattgagggt gatctaaatc cggacaactc
     5521 ggatgtcgac aaactgttca tccagttagt acaaacctat aatcagttgt ttgaagagaa
     5581 ccctataaat gcaagtggcg tggatgcgaa ggctattctt agcgcccgcc tctctaaatc
     5641 ccgacggcta gaaaacctga tcgcacaatt acccggagag aagaaaaatg ggttgttcgg
     5701 taaccttata gcgctctcac taggcctgac accaaatttt aagtcgaact tcgacttagc
     5761 tgaagatgcc aaattgcagc ttagtaagga cacgtacgat gacgatctcg acaatctact
     5821 ggcacaaatt ggagatcagt atgcggactt atttttggct gccaaaaacc ttagcgatgc
     5881 aatcctccta tctgacatac tgagagttaa tactgagatt accaaggcgc cgttatccgc
     5941 ttcaatgatc aaaaggtacg atgaacatca ccaagacttg acacttctca aggccctagt
     6001 ccgtcagcaa ctgcctgaga aatataagga aatattcttt gatcagtcga aaaacgggta
     6061 cgcaggttat attgacggcg gagcgagtca agaggaattc tacaagttta tcaaacccat
     6121 attagagaag atggatggga cggaagagtt gcttgtaaaa ctcaatcgcg aagatctact
     6181 gcgaaagcag cggactttcg acaacggtag cattccacat caaatccact taggcgaatt
     6241 gcatgctata cttagaaggc aggaggattt ttatccgttc ctcaaagaca atcgtgaaaa
     6301 gattgagaaa atcctaacct ttcgcatacc ttactatgtg ggacccctgg cccgagggaa
     6361 ctctcggttc gcatggatga caagaaagtc cgaagaaacg attactccat ggaattttga
     6421 ggaagttgtc gataaaggtg cgtcagctca atcgttcatc gagaggatga ccaactttga
     6481 caagaattta ccgaacgaaa aagtattgcc taagcacagt ttactttacg agtatttcac
     6541 agtgtacaat gaactcacga aagttaagta tgtcactgag ggcatgcgta aacccgcctt
     6601 tctaagcgga gaacagaaga aagcaatagt agatctgtta ttcaagacca accgcaaagt
     6661 gacagttaag caattgaaag aggactactt taagaaaatt gaatgcttcg attctgtcga
     6721 gatctccggg gtagaagatc gatttaatgc gtcacttggt acgtatcatg acctcctaaa
     6781 gataattaaa gataaggact tcctggataa cgaagagaat gaagatatct tagaagatat
     6841 agtgttgact cttaccctct ttgaagatcg ggaaatgatt gaggaaagac taaaaacata
     6901 cgctcacctg ttcgacgata aggttatgaa acagttaaag aggcgtcgct atacgggctg
     6961 gggacgattg tcgcggaaac ttatcaacgg gataagagac aagcaaagtg gtaaaactat
     7021 tctcgatttt ctaaagagcg acggcttcgc caataggaac tttatgcagc tgatccatga
     7081 tgactcttta accttcaaag aggatataca aaaggcacag gtttccggac aaggggactc
     7141 attgcacgaa catattgcga atcttgctgg ttcgccagcc atcaaaaagg gcatactcca
     7201 gacagtcaaa gtagtggatg agctagttaa ggtcatggga cgtcacaaac cggaaaacat
     7261 tgtaatcgag atggcacgcg aaaatcaaac gactcagaag gggcaaaaaa acagtcgaga
     7321 gcggatgaag agaatagaag agggtattaa agaactgggc agccagatct taaaggagca
     7381 tcctgtggaa aatacccaat tgcagaacga gaaactttac ctctattacc tacaaaatgg
     7441 aagggacatg tatgttgatc aggaactgga cataaaccgt ttatctgatt acgacgtcga
     7501 tgccattgta ccccaatcct ttttgaagga cgattcaatc gacaataaag tgcttacacg
     7561 ctcggataag aaccgaggga aaagtgacaa tgttccaagc gaggaagtcg taaagaaaat
     7621 gaagaactat tggcggcagc tcctaaatgc gaaactgata acgcaaagaa agttcgataa
     7681 cttaactaaa gctgagaggg gtggcttgtc tgaacttgac aaggccggat ttattaaacg
     7741 tcagctcgtg gaaacccgcc aaatcacaaa gcatgttgca cagatactag attcccgaat
     7801 gaatacgaaa tacgacgaga acgataagct gattcgggaa gtcaaagtaa tcactttaaa
     7861 gtcaaaattg gtgtcggact tcagaaagga ttttcaattc tataaagtta gggagataaa
     7921 taactaccac catgcgcacg acgcttatct taatgccgtc gtagggaccg cactcattaa
     7981 gaaatacccg aagctagaaa gtgagtttgt gtatggtgat tacaaagttt atgacgtccg
     8041 taagatgatc gcgaaaagcg aacaggagat aggcaaggct acagccaaat acttctttta
     8101 ttctaacatt atgaatttct ttaagacgga aatcactctg gcaaacggag agatacgcaa
     8161 acgaccttta attgaaacca atggggagac aggtgaaatc gtatgggata agggccggga
     8221 cttcgcgacg gtgagaaaag ttttgtccat gccccaagtc aacatagtaa agaaaactga
     8281 ggtgcagacc ggagggtttt caaaggaatc gattcttcca aaaaggaata gtgataagct
     8341 catcgctcgt aaaaaggact gggacccgaa aaagtacggt ggcttcgata gccctacagt
     8401 tgcctattct gtcctagtag tggcaaaagt tgagaaggga aaatccaaga aactgaagtc
     8461 agtcaaagaa ttattgggga taacgattat ggagcgctcg tcttttgaaa agaaccccat
     8521 cgacttcctt gaggcgaaag gttacaagga agtaaaaaag gatctcataa ttaaactacc
     8581 aaagtatagt ctgtttgagt tagaaaatgg ccgaaaacgg atgttggcta gcgccggaga
     8641 gcttcaaaag gggaacgaac tcgcactacc gtctaaatac gtgaatttcc tgtatttagc
     8701 gtcccattac gagaagttga aaggttcacc tgaagataac gaacagaagc aactttttgt
     8761 tgagcagcac aaacattatc tcgacgaaat catagagcaa atttcggaat tcagtaagag
     8821 agtcatccta gctgatgcca atctggacaa agtattaagc gcatacaaca agcacaggga
     8881 taaacccata cgtgagcagg cggaaaatat tatccatttg tttactctta ccaacctcgg
     8941 cgctccagcc gcattcaagt attttgacac aacgatagat cgcaaacgat acacttctac
     9001 caaggaggtg ctagacgcga cactgattca ccaatccatc acgggattat atgaaactcg
     9061 gatagatttg tcacagcttg ggggtgacgg atcccccaag aagaagagga aagtctcgag
     9121 cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga
     9181 tgacaagtga agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     9241 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     9301 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     9361 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     9421 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     9481 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     9541 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     9601 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     9661 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     9721 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     9781 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     9841 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     9901 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     9961 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
    10021 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
    10081 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
    10141 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
    10201 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    10261 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    10321 gct
//
