LOCUS       sequence-255052-       11912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA targeting human cGAS; Cas9 insert; Puromycin selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11912)
  AUTHORS   Barnett KC, Coronas-Serna JM, Zhou W, Ernandes MJ, Cao A, Kranzusch 
            PJ, Kagan JC
  TITLE     Phosphoinositide Interactions Position cGAS at the Plasma Membrane 
            to Ensure Efficient Distinction between Self- and Viral DNA.
  JOURNAL   Cell. 2019 Mar 7;176(6):1432-1446.e11. doi: 
            10.1016/j.cell.2019.01.049. Epub 2019 Feb 28.
  PUBMED    30827685
REFERENCE   2  (bases 1 to 11912)
  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: 
            "10.1016/j.cell.2019.01.049"; journalName: "Cell"; date: "2019-03-7-
            7"; volume: "176"; issue: "6"; pages: "1432-1446.e11"
COMMENT     Sequence Label: pRRL-gRNA(anti-cGAS)-Cas9-Puro
FEATURES             Location/Qualifiers
     source          1..11912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1575..1692
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1766..2014
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1766..1786
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1937..1956
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2036..2111
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(2123..2147)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2488..2512
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2582..2591
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2591..2656
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2663..2683
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2708..6808
                     /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             2708..6808
                     /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             6809..6856
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6866..6919
                     /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             6920..7516
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7413..7433
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7529..8117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7582..7602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8000..8011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8204..8437
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8509..8630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8546..8565)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8600..8619
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8670..8805
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8825..8844)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8826..8844)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8854..8871)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8854..8870)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8863..8885)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9012..9467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9099..9118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9309..9330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9493..9597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9598..9666,9667..10458)
                     /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(9816..9835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10629..11217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11118..11137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11371..11388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11505..11526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11541..11546,11547..11564,11565..11571)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11579..11595
                     /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     11584..11606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11603..11619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11603..11619
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11638..11658
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11640..11658
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11686..11912
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggacgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     1621 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     1681 attcaaaatt ttatcgtgta caaaaaagca ggctttaaag gaaccaattc agtcgactgg
     1741 atccggtacc aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca
     1801 tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat
     1861 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
     1921 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
     1981 ggctttatat atcttgtgga aaggacgaaa caccgagact cggtgggatc catcggtttt
     2041 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     2101 cgagtcggtg ctttttttct agacccagct ttcttgtaca aagttggcat tacgattacg
     2161 cgtaggaaca gagaaacagg agaatatggg ccaaacagga tatctgtggt aagcagttcc
     2221 tgccccggct cagggccaag aacagttgga acagcagaat atgggccaaa caggatatct
     2281 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     2341 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     2401 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     2461 tctgctcccc gagctctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     2521 acgccatcca cgctgttttg acttccatag aaggatctcg agcggccgcg ctagcggatc
     2581 cgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
     2641 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
     2701 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     2761 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     2821 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     2881 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     2941 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     3001 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     3061 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     3121 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     3181 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     3241 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     3301 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     3361 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     3421 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     3481 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     3541 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     3601 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     3661 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     3721 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     3781 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     3841 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     3901 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     3961 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     4021 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     4081 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     4141 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     4201 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     4261 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     4321 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     4381 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     4441 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     4501 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     4561 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     4621 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     4681 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     4741 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     4801 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     4861 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     4921 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     4981 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     5041 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     5101 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     5161 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     5221 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     5281 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     5341 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     5401 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     5461 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     5521 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     5581 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     5641 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     5701 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     5761 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     5821 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     5881 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     5941 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     6001 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     6061 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     6121 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     6181 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     6241 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     6301 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     6361 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     6421 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     6481 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     6541 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     6601 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     6661 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     6721 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     6781 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     6841 ggcaaaaaag aaaaagggat ccggtgaggg cagaggaagt cttctaacat gcggtgacgt
     6901 ggaggagaat ccgggcccta ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     6961 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     7021 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     7081 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     7141 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     7201 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     7261 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     7321 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     7381 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     7441 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     7501 caagcccggt gcctgaacta gtgtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     7561 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     7621 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     7681 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     7741 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     7801 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     7861 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     7921 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     7981 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     8041 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     8101 ttgggccgcc tccccgcctg gaattcgagc tcggtacctt taagaccaat gacttacaag
     8161 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     8221 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     8281 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     8341 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     8401 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     8461 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     8521 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     8581 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     8641 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     8701 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     8761 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     8821 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     8881 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     8941 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     9001 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     9061 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     9121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     9181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     9241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     9301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     9361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     9421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     9481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     9541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     9601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     9661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     9721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     9781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     9841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     9901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     9961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
    10021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
    10081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
    10141 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
    10201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
    10261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
    10321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
    10381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
    10441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
    10501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
    10561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
    10621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
    10681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
    10741 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
    10801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
    10861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
    10921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
    10981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
    11041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
    11101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
    11161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    11221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
    11281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    11341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
    11401 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
    11461 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
    11521 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
    11581 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
    11641 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
    11701 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
    11761 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
    11821 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
    11881 ttgtatttaa gtgcctaact cgatacataa ac
//
