LOCUS       sequence-168914-        8832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DD-SpCas9 in mammalian cells and has a cloning site for an
            sgRNA. The FKBP12 L106P destabilization domain allows for inducible 
            stabilization of Cas9 through the addition of Shield-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8832)
  AUTHORS   Geisinger JM, Turan S, Hernandez S, Spector LP, Calos MP
  TITLE     In vivo blunt-end cloning through CRISPR/Cas9-facilitated 
            non-homologous end-joining.
  JOURNAL   Nucleic Acids Res. 2016 Jan 13. pii: gkv1542.
  PUBMED    26762978
REFERENCE   2  (bases 1 to 8832)
  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: "Nucleic 
            Acids Res. 2016 Jan 13. pii: gkv1542."
COMMENT     Sequence Label: pX330-BsaIx2-DD-Cas9
FEATURES             Location/Qualifiers
     source          1..8832
                     /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        267..342
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        439..724
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        726..1003
                     /label=chicken beta-actin promoter
     intron          1004..1232
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1244..1253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1250..1252,1253..1573)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             1580..1645
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1652..1672
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1697..5797
                     /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             1697..5797
                     /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             5798..5845
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5873..5890)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5879..6086
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6095..6235
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6095..6224
                     /label=AAV2 ITR
     rep_origin      6310..6765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6397..6416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6607..6628
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6782..6801)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6901..6923
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6961..6979)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7047..7151
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7152..7220,7221..8012)
                     /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(7370..7389)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8183..8771
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8672..8691
                     /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 ggagaccgag gtctctgttt tagagctaga aatagcaagt taaaataagg
      301 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt tttagagcta
      361 gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat tctgcagaca
      421 aatggctcta gaggtacccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      481 caacgacccc cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca
      541 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      601 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta
      661 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      721 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      781 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      841 gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
      901 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      961 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgacgct
     1021 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     1081 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     1141 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
     1201 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tgggagtgca
     1261 ggtggaaacc atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgtgt
     1321 ggtgcactac accgggatgc ttgaagatgg aaagaaagtc gattcctccc gggacagaaa
     1381 caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt
     1441 tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg
     1501 tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct
     1561 tctaaaaccg gaagaattcg actataagga ccacgacgga gactacaagg atcatgatat
     1621 tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca
     1681 cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt
     1741 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     1801 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
     1861 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     1921 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     1981 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     2041 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     2101 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     2161 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     2221 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     2281 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     2341 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     2401 cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     2461 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     2521 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     2581 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     2641 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     2701 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     2761 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     2821 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     2881 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     2941 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     3001 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     3061 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     3121 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     3181 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     3241 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     3301 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     3361 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     3421 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     3481 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     3541 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     3601 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     3661 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     3721 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     3781 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     3841 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     3901 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     3961 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     4021 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     4081 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     4141 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     4201 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     4261 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     4321 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     4381 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     4441 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     4501 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     4561 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     4621 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     4681 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     4741 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     4801 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     4861 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     4921 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     4981 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     5041 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     5101 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     5161 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     5221 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     5281 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     5341 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     5401 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     5461 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     5521 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     5581 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     5641 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     5701 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     5761 cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa
     5821 ggccggccag gcaaaaaaga aaaagtaaga attcctagag ctcgctgatc agcctcgact
     5881 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5941 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     6001 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     6061 gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat ggagttggcc
     6121 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     6181 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     6241 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     6301 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6361 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6421 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6481 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     6541 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6601 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     6661 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6721 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     6781 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6841 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6901 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6961 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     7021 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7081 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7141 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7201 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7261 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7321 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7381 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7441 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7501 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7561 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7621 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7681 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7741 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7801 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7861 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7921 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7981 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     8041 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     8101 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     8161 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     8221 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     8281 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     8341 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     8401 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     8461 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     8521 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8581 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8641 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8701 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8761 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8821 ttgctcacat gt
//
