LOCUS       sequence-319242-        9284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription of p53 guide RNA and Cas9 expression for CRISPR/Cas9 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9284)
  AUTHORS   Mohr L, Toufektchan E, von Morgen P, Chu K, Kapoor A, Maciejowski J
  TITLE     ER-directed TREX1 limits cGAS activation at micronuclei.
  JOURNAL   Mol Cell. 2021 Jan 12. pii: S1097-2765(20)30958-8. doi: 
            10.1016/j.molcel.2020.12.037.
  PUBMED    33476576
REFERENCE   2  (bases 1 to 9284)
  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: "Mol Cell. 
            2021 Jan 12. pii: S1097-2765(20)30958-8. doi: 
            10.1016/j.molcel.2020.12.037."
COMMENT     Sequence Label: pU6-p53 gRNA-Cas9-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..9284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..255
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7..27
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     178..197
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        275..350
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        447..732
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        734..1011
                     /label=chicken beta-actin promoter
     intron          1012..1240
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1252..1261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1261..1326
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1333..1353
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1378..5478
                     /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             1378..5478
                     /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             5479..5526
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5530..5583
                     /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             5590..6300
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5735..5753)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(6325..6342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6331..6538
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6547..6687
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6547..6676
                     /label=AAV2 ITR
     rep_origin      6762..7217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6849..6868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7059..7080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7234..7253)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7353..7375
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7413..7431)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7499..7603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7604..7672,7673..8464)
                     /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(7822..7841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8635..9223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9124..9143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccccacgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
       61 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
      121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
      181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      241 aaaggacgaa acaccggcag ctacggtttc cgtcgtttta gagctagaaa tagcaagtta
      301 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt
      361 tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc
      421 tgcagacaaa tggctctaga ggtacccgtt acataactta cggtaaatgg cccgcctggc
      481 tgaccgccca acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat
      541 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      601 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt
      661 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      721 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
      781 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
      841 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
      901 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
      961 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     1021 gcgacgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
     1081 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     1141 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
     1201 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg
     1261 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
     1321 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
     1381 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
     1441 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     1501 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     1561 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     1621 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     1681 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     1741 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     1801 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     1861 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     1921 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     1981 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     2041 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     2101 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     2161 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     2221 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     2281 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     2341 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     2401 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     2461 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     2521 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     2581 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     2641 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     2701 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     2761 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     2821 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     2881 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     2941 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     3001 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     3061 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     3121 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     3181 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     3241 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     3301 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     3361 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     3421 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     3481 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     3541 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     3601 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     3661 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     3721 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     3781 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     3841 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     3901 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     3961 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     4021 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     4081 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     4141 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     4201 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     4261 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     4321 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     4381 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     4441 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     4501 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     4561 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     4621 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     4681 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     4741 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     4801 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     4861 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     4921 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     4981 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     5041 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     5101 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     5161 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     5221 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     5281 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     5341 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     5401 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     5461 tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     5521 aaaaagcttg agggcagagg aagtctgcta acatgcggtg acgtggagga gaatcccggc
     5581 cctgctagca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     5641 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     5701 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     5761 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     5821 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     5881 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     5941 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctcagac
     6001 ggccccgtaa tgcagaagaa aaccatgggc tgggaggcct cctccgagcg gatgtacccc
     6061 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     6121 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     6181 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     6241 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     6301 gaattcctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     6361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     6481 gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc
     6541 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     6601 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     6661 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     6721 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     6781 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6841 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6901 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6961 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     7021 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     7081 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     7141 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     7201 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7261 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     7321 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     7381 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     7441 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7501 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7561 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7621 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7681 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7741 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7801 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7861 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7921 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7981 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     8041 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8101 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     8161 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8221 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8281 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     8341 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8401 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8461 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8521 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8581 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8641 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8701 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8761 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     8821 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8881 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8941 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     9001 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     9061 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     9121 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     9181 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9241 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
