LOCUS       sequence-366457-       11911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA targeting mouse STING (with Cas9 insert).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11911)
  AUTHORS   Mosallanejad K, Kennedy SN, Bahleda KM, Slavik KM, Zhou W, Govande 
            AA, Hancks DC, Kranzusch PJ, Kagan JC
  TITLE     Species-specific self-DNA detection mechanisms by mammalian cyclic 
            GMP-AMP synthases.
  JOURNAL   Sci Immunol. 2023 Jan 20;8(79):eabp9765. doi: 
            10.1126/sciimmunol.abp9765. Epub 2023 Jan 20.
  PUBMED    36662885
REFERENCE   2  (bases 1 to 11911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci 
            Immunol."; date: "2023-01-20"; volume: "8(79):eabp9765. doi"; pages:
            " 10.1126/sciimmunol.abp9765. Epub 2023 Jan 20"
COMMENT     Sequence Label: pRRL-mouse STING #1-gRNA-Cas9-Puro
FEATURES             Location/Qualifiers
     source          1..11911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..43
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(58..63,64..81,82..88)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    96..112
                     /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     101..123
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     120..136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     120..136
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     155..175
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        157..175
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        203..429
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             430..610
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    657..782
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1275..1508
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1693..1737
                     /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    2004..2121
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2195..2443
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2195..2215
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2366..2385
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2464..2539
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(2551..2575)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2916..2940
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3010..3019
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3019..3084
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3091..3111
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3136..7236
                     /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             3136..7236
                     /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             7237..7284
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7294..7347
                     /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             7348..7944
                     /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     7841..7861
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7957..8545
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8010..8030)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8428..8439)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8632..8865
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8937..9058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8974..8993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9028..9047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9098..9233
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9253..9272)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9254..9272)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9282..9299)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9282..9298)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9291..9313)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9440..9895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9527..9546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9737..9758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9921..10025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10026..10094,10095..10886)
                     /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(10244..10263)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11057..11645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11546..11565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11799..11816
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
       61 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      121 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
      181 aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg
      241 gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg
      301 gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg
      361 attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg cctaactcga
      421 tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
      481 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
      541 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
      601 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
      661 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
      721 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
      781 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
      841 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
      901 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
      961 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     1021 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     1081 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggacgctgat cttcagacct
     1141 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     1201 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     1261 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     1321 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     1381 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     1441 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     1501 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     1561 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     1621 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     1681 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     1741 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     1801 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     1861 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     1921 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     1981 ggatctcgac ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca
     2041 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     2101 attacaaaaa ttcaaaattt tatcgtgtac aaaaaagcag gctttaaagg aaccaattca
     2161 gtcgactgga tccggtacca aggtcgggca ggaagagggc ctatttccca tgattccttc
     2221 atatttgcat atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa
     2281 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
     2341 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
     2401 cgatttcttg gctttatata tcttgtggaa aggacgaaac accgcgaggc taggtgaagt
     2461 gctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     2521 agtggcaccg agtcggtgct ttttttctag acccagcttt cttgtacaaa gttggcatta
     2581 cgattacgcg taggaacaga gaaacaggag aatatgggcc aaacaggata tctgtggtaa
     2641 gcagttcctg ccccggctca gggccaagaa cagttggaac agcagaatat gggccaaaca
     2701 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     2761 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     2821 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     2881 cgcgcgcttc tgctccccga gctctatata agcagagctc gtttagtgaa ccgtcagatc
     2941 gcctggagac gccatccacg ctgttttgac ttccatagaa ggatctcgag cggccgcgct
     3001 agcggatccg ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt
     3061 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
     3121 ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
     3181 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     3241 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     3301 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     3361 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     3421 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     3481 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     3541 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     3601 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     3661 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     3721 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     3781 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     3841 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     3901 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     3961 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     4021 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     4081 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     4141 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     4201 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     4261 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     4321 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     4381 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     4441 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     4501 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     4561 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     4621 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     4681 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     4741 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     4801 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     4861 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     4921 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     4981 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     5041 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     5101 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     5161 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     5221 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     5281 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     5341 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     5401 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     5461 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     5521 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     5581 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     5641 gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     5701 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     5761 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     5821 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     5881 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     5941 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     6001 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     6061 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     6121 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     6181 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     6241 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     6301 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     6361 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     6421 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     6481 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     6541 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     6601 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     6661 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     6721 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     6781 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     6841 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     6901 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     6961 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     7021 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     7081 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     7141 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     7201 gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag
     7261 gccggccagg caaaaaagaa aaagggatcc ggtgagggca gaggaagtct tctaacatgc
     7321 ggtgacgtgg aggagaatcc gggccctacc gagtacaagc ccacggtgcg cctcgccacc
     7381 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     7441 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     7501 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     7561 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     7621 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     7681 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     7741 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     7801 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     7861 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     7921 atgacccgca agcccggtgc ctgaactagt gtcgacaatc aacctctgga ttacaaaatt
     7981 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     8041 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     8101 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     8161 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     8221 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     8281 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     8341 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     8401 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     8461 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     8521 atctcccttt gggccgcctc cccgcctgga attcgagctc ggtaccttta agaccaatga
     8581 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     8641 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     8701 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     8761 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     8821 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     8881 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     8941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     9001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     9061 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     9121 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     9181 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac
     9241 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     9301 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     9361 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     9421 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     9481 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     9541 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     9601 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     9661 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     9721 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     9781 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     9841 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     9901 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     9961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
    10021 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
    10081 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
    10141 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
    10201 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
    10261 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
    10321 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
    10381 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
    10441 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
    10501 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
    10561 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
    10621 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
    10681 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
    10741 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
    10801 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
    10861 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
    10921 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
    10981 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
    11041 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
    11101 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
    11161 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
    11221 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
    11281 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
    11341 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
    11401 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
    11461 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    11521 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    11581 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    11641 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
    11701 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
    11761 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
    11821 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
    11881 gctggcacga caggtttccc gactggaaag c
//
