LOCUS       sequence-240504-        8487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses WT spCas9 and a chimeric gRNA targeting human LMNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  AUTHORS   Pinder J, Salsman J, Dellaire G
  TITLE     Nuclear domain 'knock-in' screen for the evaluation and 
            identification of small molecule enhancers of CRISPR-based genome 
            editing.
  JOURNAL   Nucleic Acids Res. 2015 Oct 30;43(19):9379-92. doi: 
            10.1093/nar/gkv993. Epub 2015 Oct 1.
  PUBMED    26429972
REFERENCE   2  (bases 1 to 8487)
  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"; doi: 
            "10.1093/nar/gkv993"; journalName: "Nucleic Acids Res"; date: 
            "2015-10-30- 30"; volume: "43"; issue: "19"; pages: "9379-92"
COMMENT     Sequence Label: pX330-LMNA-gRNA1
FEATURES             Location/Qualifiers
     source          1..8487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          142..369
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      381..390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             390..455
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             462..482
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             507..4607
                     /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             507..4607
                     /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             4608..4655
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4683..4700)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4689..4896
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4905..5045
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4905..5034
                     /label=AAV2 ITR
     rep_origin      5120..5575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5207..5226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5417..5438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5592..5611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5711..5733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5771..5789)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5857..5961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5962..6030,6031..6822)
                     /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(6180..6199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6993..7581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7482..7501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7643..7891
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7643..7663
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7814..7833
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7913..7988
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8085..8370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8372..8487,1..141)
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
ORIGIN
        1 ggggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
       61 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
      121 aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc
      181 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
      241 gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag
      301 ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact
      361 ttttttcagg ttggaccggt gccaccatgg actataagga ccacgacgga gactacaagg
      421 atcatgatat tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg
      481 tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca
      541 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
      601 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
      661 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
      721 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
      781 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
      841 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
      901 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
      961 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     1021 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     1081 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     1141 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     1201 agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca
     1261 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     1321 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     1381 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     1441 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     1501 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     1561 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     1621 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     1681 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     1741 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     1801 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     1861 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     1921 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
     1981 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     2041 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
     2101 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
     2161 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     2221 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     2281 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     2341 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     2401 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     2461 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     2521 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     2581 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     2641 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     2701 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     2761 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     2821 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     2881 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     2941 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     3001 tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg
     3061 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
     3121 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
     3181 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     3241 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     3301 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
     3361 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     3421 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     3481 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     3541 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     3601 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     3661 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
     3721 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
     3781 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     3841 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     3901 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     3961 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     4021 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     4081 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     4141 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     4201 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     4261 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
     4321 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
     4381 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
     4441 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     4501 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     4561 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg
     4621 ccacgaaaaa ggccggccag gcaaaaaaga aaaagtaaga attcctagag ctcgctgatc
     4681 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4741 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4801 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4861 ggaggattgg gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat
     4921 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4981 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     5041 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     5101 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5161 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     5221 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5281 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     5341 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5401 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     5461 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     5521 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     5581 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5641 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5701 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5761 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5821 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5881 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5941 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6001 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6061 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6121 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6181 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6241 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6301 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6361 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6421 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6481 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6541 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6601 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6661 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     6721 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6781 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6841 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6901 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6961 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7021 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7081 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7141 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7201 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7261 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7321 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7381 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7441 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7501 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7561 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7621 tgctggcctt ttgctcacat gtgagggcct atttcccatg attccttcat atttgcatat
     7681 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt
     7741 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
     7801 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
     7861 tttatatatc ttgtggaaag gacgaaacac cgggttggca gcgctgcccg cggttttaga
     7921 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     7981 gtcggtgctt ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt
     8041 tagcgcgtgc gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg
     8101 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg
     8161 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     8221 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     8281 tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac
     8341 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     8401 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     8461 tgtgcagcga tgggggcggg ggggggg
//
