LOCUS       sequence-366450-       11911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA targeting mouse cGAS (with Cas9 insert).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11911)
  AUTHORS   Mosallanejad K, Kennedy SN, Bahleda KM, Slavik KM, Zhou W, Govande 
            AA, Hancks DC, Kranzusch PJ, Kagan JC
  TITLE     Species-specific self-DNA detection mechanisms by mammalian cyclic 
            GMP-AMP synthases.
  JOURNAL   Sci Immunol. 2023 Jan 20;8(79):eabp9765. doi: 
            10.1126/sciimmunol.abp9765. Epub 2023 Jan 20.
  PUBMED    36662885
REFERENCE   2  (bases 1 to 11911)
  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: "Sci 
            Immunol."; date: "2023-01-20"; volume: "8(79):eabp9765. doi"; pages:
            " 10.1126/sciimmunol.abp9765. Epub 2023 Jan 20"
COMMENT     Sequence Label: pRRL-mouse cGAS #1-gRNA-Cas9-Puro
FEATURES             Location/Qualifiers
     source          1..11911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     390..410
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    506..1094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(559..579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(977..988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1181..1414
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1486..1607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1523..1542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1577..1596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1647..1782
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1802..1821)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1803..1821)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1831..1848)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1831..1847)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1840..1862)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1989..2444
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2076..2095)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2286..2307
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2470..2574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2575..2643,2644..3435)
                     /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(2793..2812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3606..4194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4095..4114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4348..4365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4482..4503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4518..4523,4524..4541,4542..4548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4556..4572
                     /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)."
     primer_bind     4561..4583
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4580..4596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4580..4596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4615..4635
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4617..4635
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4663..4889
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4890..5070
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5117..5242
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5735..5968
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6153..6197
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6464..6581
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6655..6903
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6655..6675
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6826..6845
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6924..6999
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(7011..7035)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     7376..7400
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      7470..7479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7479..7544
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7551..7571
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7596..11696
                     /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             7596..11696
                     /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             11697..11744
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             11754..11807
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(11808..11911,1..493)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
ORIGIN
        1 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
       61 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
      121 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
      181 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
      241 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
      301 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
      361 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
      421 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
      481 gcccggtgcc tgaactagtg tcgacaatca acctctggat tacaaaattt gtgaaagatt
      541 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
      601 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
      661 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
      721 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
      781 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
      841 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
      901 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
      961 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     1021 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     1081 ggccgcctcc ccgcctggaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca
     1141 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     1201 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     1261 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     1321 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     1381 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     1441 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     1501 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1561 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     1621 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1681 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     1741 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     1801 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     1861 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     1921 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1981 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2041 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2101 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2161 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2221 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2281 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2341 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2401 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     2461 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2521 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2581 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2641 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2701 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2761 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2821 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2881 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2941 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3001 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3061 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3121 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3181 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3241 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3301 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3361 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3421 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3481 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3541 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3601 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3661 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3721 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3781 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3841 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3901 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3961 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4021 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4081 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4141 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4201 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4261 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4321 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     4381 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     4441 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     4501 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     4561 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     4621 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     4681 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     4741 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     4801 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     4861 tatttaagtg cctaactcga tacataaacg ggtctctctg gttagaccag atctgagcct
     4921 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     4981 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     5041 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     5101 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     5161 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     5221 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     5281 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     5341 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     5401 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     5461 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     5521 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     5581 ggacgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     5641 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     5701 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     5761 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     5821 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     5881 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     5941 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     6001 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     6061 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     6121 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     6181 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     6241 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     6301 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     6361 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     6421 gatccattcg attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg
     6481 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     6541 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tatcgtgtac aaaaaagcag
     6601 gctttaaagg aaccaattca gtcgactgga tccggtacca aggtcgggca ggaagagggc
     6661 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
     6721 agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
     6781 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
     6841 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
     6901 accgaggcgc ggaaagtcgt aaggttttag agctagaaat agcaagttaa aataaggcta
     6961 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttctag acccagcttt
     7021 cttgtacaaa gttggcatta cgattacgcg taggaacaga gaaacaggag aatatgggcc
     7081 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagttggaac
     7141 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     7201 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     7261 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     7321 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctctatata agcagagctc
     7381 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ttccatagaa
     7441 ggatctcgag cggccgcgct agcggatccg ccaccatgga ctataaggac cacgacggag
     7501 actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga
     7561 agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg
     7621 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     7681 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     7741 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     7801 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     7861 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     7921 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     7981 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     8041 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     8101 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     8161 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     8221 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     8281 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     8341 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     8401 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     8461 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     8521 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     8581 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     8641 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     8701 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     8761 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     8821 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     8881 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     8941 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     9001 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     9061 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     9121 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     9181 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     9241 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     9301 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     9361 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     9421 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     9481 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     9541 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     9601 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     9661 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     9721 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     9781 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     9841 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     9901 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     9961 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
    10021 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
    10081 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
    10141 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
    10201 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
    10261 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
    10321 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
    10381 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
    10441 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
    10501 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
    10561 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
    10621 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
    10681 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
    10741 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
    10801 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
    10861 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
    10921 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
    10981 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
    11041 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
    11101 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
    11161 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
    11221 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
    11281 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
    11341 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
    11401 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
    11461 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
    11521 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
    11581 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
    11641 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa
    11701 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagggatcc ggtgagggca
    11761 gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc gggccctacc gagtacaagc
    11821 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
    11881 cgttcgccga ctaccccgcc acgcgccaca c
//
