LOCUS       sequence-202376-       13837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 lentivector targeting human cGAS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13837)
  AUTHORS   Gentili M, Kowal J, Tkach M, Satoh T, Lahaye X, Conrad C, Boyron M, 
            Lombard B, Durand S, Kroemer G, Loew D, Dalod M, Thery C, Manel N
  TITLE     Transmission of innate immune signaling by packaging of cGAMP in 
            viral particles.
  JOURNAL   Science. 2015 Sep 11;349(6253):1232-6. doi: 10.1126/science.aab3628.
            Epub 2015 Jul 30.
  PUBMED    26229115
REFERENCE   2  (bases 1 to 13837)
  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: 
            "10.1126/science.aab3628"; journalName: "Science"; date: 
            "2015-09-11- 11"; volume: "349"; issue: "6253"; pages: "1232-6"
COMMENT     Sequence Label: pLCRISPR-CMV-cGAS3
FEATURES             Location/Qualifiers
     source          1..13837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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"
     promoter        2397..2645
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2397..2417
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2568..2587
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2667..2742
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2776..2893
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2988..3291
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3292..3495
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3549..3558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3558..3623
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3630..3650
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3675..7775
                     /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             3675..7775
                     /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             7785..7832
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7842..7898
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7905..8504
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7905..7924)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8401..8421
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8529..9117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8582..8602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9000..9011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9120..9136)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9121..9137)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9642..9822
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9848..9865)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9854..10078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10124..10552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10211..10230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10421..10442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10561..10581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10566..10895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10746..10881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10808..10827
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10943..10990
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11009..11383
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11513..11634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11550..11569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11604..11623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11683..11699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11683..11699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11696..11718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11707..11723
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(11731..11737,11738..11755,11756..11761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11776..11797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11914..11931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12085..12673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12165..12184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12844..13635,13636..13704))
                     /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     13467..13486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13705..13809)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgagg
     2401 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     2461 ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     2521 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     2581 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     2641 acaccgggga tcccgacttc ctggcggttt tagagctaga aatagcaagt taaaataagg
     2701 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttta gacaaatggc
     2761 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     2821 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     2881 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt taattaatag
     2941 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3001 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3061 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3121 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     3181 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     3241 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3301 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3361 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3421 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3481 tatataagca gagctggttt agtgaaccgt cagatcccgc gggctagcgg atccccaccg
     3541 gtcgtacggc caccatggac tataaggacc acgacggaga ctacaaggat catgatattg
     3601 attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg
     3661 gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
     3721 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     3781 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     3841 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     3901 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     3961 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     4021 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     4081 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     4141 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     4201 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     4261 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     4321 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     4381 tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     4441 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     4501 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     4561 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     4621 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     4681 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     4741 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     4801 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     4861 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     4921 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     4981 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     5041 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     5101 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     5161 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     5221 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     5281 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     5341 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     5401 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     5461 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     5521 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     5581 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     5641 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     5701 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     5761 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     5821 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     5881 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     5941 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     6001 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     6061 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     6121 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     6181 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     6241 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     6301 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     6361 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     6421 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     6481 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     6541 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     6601 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     6661 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     6721 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     6781 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     6841 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     6901 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     6961 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     7021 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     7081 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     7141 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     7201 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     7261 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     7321 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     7381 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     7441 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     7501 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     7561 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     7621 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     7681 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     7741 agacacggat cgacctgtct cagctgggag gcgacgcggc cgccaagcgt cctgctgcta
     7801 ctaagaaagc tggtcaagct aagaaaaaga aaggaagcgg agctactaac ttcagcctgc
     7861 tgaagcaggc tggagacgtg gaggagaacc ctggacctgg atccatgacc gagtacaagc
     7921 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     7981 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     8041 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     8101 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     8161 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     8221 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     8281 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     8341 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     8401 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     8461 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctaagaattc gatatcaagc
     8521 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     8581 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     8641 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     8701 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     8761 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     8821 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     8881 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     8941 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     9001 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     9061 cgcatcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     9121 gataccgtcg acctcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     9181 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     9241 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     9301 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaagt caagatatcc
     9361 ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag
     9421 ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     9481 aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc
     9541 tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag
     9601 catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc
     9661 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     9721 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     9781 gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc
     9841 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     9901 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     9961 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
    10021 aagggggagg attgggaaga taatagcagg catgctgggg atgcggtggg ctctatggct
    10081 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
    10141 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
    10201 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
    10261 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    10321 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    10381 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    10441 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
    10501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
    10561 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
    10621 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
    10681 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
    10741 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
    10801 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
    10861 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
    10921 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
    10981 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct
    11041 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
    11101 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
    11161 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
    11221 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
    11281 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
    11341 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
    11401 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
    11461 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
    11521 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
    11581 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
    11641 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
    11701 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
    11761 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
    11821 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
    11881 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
    11941 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
    12001 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    12061 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    12121 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    12181 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    12241 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    12301 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    12361 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    12421 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    12481 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
    12541 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    12601 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    12661 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
    12721 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
    12781 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
    12841 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
    12901 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
    12961 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
    13021 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
    13081 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
    13141 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
    13201 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
    13261 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
    13321 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
    13381 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
    13441 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
    13501 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
    13561 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
    13621 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
    13681 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
    13741 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
    13801 ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
