LOCUS       sequence-193934-       13839 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 13839)
  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 13839)
  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..13839
                     /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        2989..3292
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3293..3496
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3551..3560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3560..3625
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3632..3652
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3677..7777
                     /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             3677..7777
                     /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             7787..7834
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7844..7900
                     /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             7907..8506
                     /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(7907..7926)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8403..8423
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8531..9119
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8584..8604)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9002..9013)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9122..9138)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9123..9139)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9644..9824
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9850..9867)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9856..10080
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10126..10554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10213..10232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10423..10444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10563..10583)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10568..10897
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10748..10883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10810..10829
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10945..10992
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11011..11385
                     /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    11515..11636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11552..11571)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11606..11625
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11685..11701)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11685..11701)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11698..11720)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11709..11725
                     /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(11733..11739,11740..11757,11758..11763))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11778..11799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11916..11933)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12087..12675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12167..12186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12846..13637,13638..13706))
                     /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     13469..13488
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13707..13811)
                     /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 taattaaata
     2941 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     3001 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     3061 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     3121 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     3181 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     3241 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     3301 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     3361 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     3421 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     3481 ctatataagc agagctggtt tagtgaaccg tcagatcccg cgggctagcg gatccccacc
     3541 ggtccgtacg gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat
     3601 tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca
     3661 cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt
     3721 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
     3781 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
     3841 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
     3901 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
     3961 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
     4021 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
     4081 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
     4141 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
     4201 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
     4261 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
     4321 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
     4381 cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     4441 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     4501 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     4561 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     4621 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     4681 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     4741 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     4801 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     4861 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     4921 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     4981 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     5041 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     5101 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     5161 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     5221 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     5281 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     5341 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     5401 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     5461 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     5521 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     5581 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     5641 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     5701 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     5761 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     5821 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     5881 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     5941 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     6001 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     6061 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     6121 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     6181 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     6241 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     6301 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     6361 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     6421 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     6481 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     6541 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     6601 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     6661 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     6721 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     6781 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     6841 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     6901 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     6961 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     7021 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     7081 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     7141 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     7201 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     7261 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     7321 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     7381 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     7441 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     7501 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     7561 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     7621 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     7681 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     7741 cgagacacgg atcgacctgt ctcagctggg aggcgacgcg gccgccaagc gtcctgctgc
     7801 tactaagaaa gctggtcaag ctaagaaaaa gaaaggaagc ggagctacta acttcagcct
     7861 gctgaagcag gctggagacg tggaggagaa ccctggacct ggatccatga ccgagtacaa
     7921 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     7981 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     8041 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     8101 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     8161 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     8221 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     8281 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     8341 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     8401 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     8461 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctaagaat tcgatatcaa
     8521 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     8581 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     8641 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     8701 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     8761 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     8821 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     8881 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     8941 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     9001 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     9061 tccgcatctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     9121 tcgataccgt cgacctcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     9181 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     9241 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     9301 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gtcaagatat
     9361 ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga actacacacc
     9421 agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     9481 gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag
     9541 cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct
     9601 agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga
     9661 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     9721 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     9781 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc
     9841 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     9901 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     9961 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
    10021 gcaaggggga ggattgggaa gataatagca ggcatgctgg ggatgcggtg ggctctatgg
    10081 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
    10141 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
    10201 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
    10261 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
    10321 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
    10381 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
    10441 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
    10501 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
    10561 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
    10621 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
    10681 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
    10741 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
    10801 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
    10861 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
    10921 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
    10981 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
    11041 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
    11101 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
    11161 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
    11221 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
    11281 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
    11341 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
    11401 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
    11461 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
    11521 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
    11581 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
    11641 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
    11701 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
    11761 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
    11821 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
    11881 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
    11941 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
    12001 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
    12061 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    12121 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    12181 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    12241 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    12301 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    12361 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    12421 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    12481 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
    12541 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    12601 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    12661 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    12721 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
    12781 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
    12841 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
    12901 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
    12961 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
    13021 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
    13081 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
    13141 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
    13201 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
    13261 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
    13321 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
    13381 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
    13441 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
    13501 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
    13561 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
    13621 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
    13681 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
    13741 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
    13801 ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
