LOCUS       sequence-366478-       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 #2-gRNA-Cas9-Puro
FEATURES             Location/Qualifiers
     source          1..11911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        696..922
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             923..1103
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1150..1275
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1768..2001
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2186..2230
                     /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    2497..2614
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2688..2936
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2688..2708
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2859..2878
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2957..3032
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(3044..3068)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     3409..3433
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3503..3512
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3512..3577
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3584..3604
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3629..7729
                     /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             3629..7729
                     /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             7730..7777
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7787..7840
                     /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             7841..8437
                     /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     8334..8354
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8450..9038
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8503..8523)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8921..8932)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9125..9358
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9430..9551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9467..9486)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9521..9540
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9591..9726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9746..9765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9747..9765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9775..9792)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9775..9791)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9784..9806)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9933..10388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10020..10039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10230..10251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10414..10518
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10519..10587,10588..11379)
                     /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(10737..10756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11550..11911,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      661 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
      721 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
      781 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
      841 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
      901 gtgcctaact cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
      961 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     1021 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     1081 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     1141 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     1201 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     1261 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     1321 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     1381 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     1441 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     1501 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     1561 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggacgct
     1621 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     1681 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     1741 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     1801 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     1861 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     1921 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     1981 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     2041 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     2101 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     2161 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     2221 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     2281 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     2341 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     2401 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     2461 tcgattagtg aacggatctc gacggtatcg gttaactttt aaaagaaaag gggggattgg
     2521 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     2581 attacaaaaa caaattacaa aaattcaaaa ttttatcgtg tacaaaaaag caggctttaa
     2641 aggaaccaat tcagtcgact ggatccggta ccaaggtcgg gcaggaagag ggcctatttc
     2701 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attagaatta
     2761 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
     2821 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
     2881 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgctg
     2941 gccaccagaa agatgagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     3001 atcaacttga aaaagtggca ccgagtcggt gctttttttc tagacccagc tttcttgtac
     3061 aaagttggca ttacgattac gcgtaggaac agagaaacag gagaatatgg gccaaacagg
     3121 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagttgg aacagcagaa
     3181 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     3241 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     3301 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     3361 tctcgcttct gttcgcgcgc ttctgctccc cgagctctat ataagcagag ctcgtttagt
     3421 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacttccata gaaggatctc
     3481 gagcggccgc gctagcggat ccgccaccat ggactataag gaccacgacg gagactacaa
     3541 ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa
     3601 ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg
     3661 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
     3721 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
     3781 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
     3841 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
     3901 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
     3961 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
     4021 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
     4081 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
     4141 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
     4201 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
     4261 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
     4321 gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt
     4381 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
     4441 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
     4501 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
     4561 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
     4621 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
     4681 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
     4741 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
     4801 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
     4861 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
     4921 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
     4981 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
     5041 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
     5101 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
     5161 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
     5221 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
     5281 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
     5341 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
     5401 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
     5461 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
     5521 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
     5581 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
     5641 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
     5701 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
     5761 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
     5821 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
     5881 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
     5941 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
     6001 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
     6061 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
     6121 gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat
     6181 cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc
     6241 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
     6301 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
     6361 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
     6421 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
     6481 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
     6541 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
     6601 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
     6661 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
     6721 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
     6781 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
     6841 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
     6901 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
     6961 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
     7021 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
     7081 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
     7141 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
     7201 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
     7261 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
     7321 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
     7381 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
     7441 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
     7501 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
     7561 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     7621 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     7681 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc
     7741 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggga tccggtgagg gcagaggaag
     7801 tcttctaaca tgcggtgacg tggaggagaa tccgggccct accgagtaca agcccacggt
     7861 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     7921 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     7981 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     8041 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     8101 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     8161 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     8221 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     8281 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     8341 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     8401 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaact agtgtcgaca atcaacctct
     8461 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     8521 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     8581 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     8641 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     8701 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     8761 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     8821 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     8881 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     8941 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     9001 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacct
     9061 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     9121 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     9181 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     9241 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     9301 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     9361 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     9421 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     9481 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     9541 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc
     9601 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     9661 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     9721 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     9781 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     9841 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     9901 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     9961 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
    10021 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
    10081 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
    10141 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
    10201 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
    10261 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
    10321 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
    10381 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
    10441 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
    10501 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
    10561 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
    10621 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
    10681 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
    10741 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
    10801 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
    10861 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
    10921 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
    10981 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
    11041 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
    11101 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
    11161 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
    11221 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
    11281 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
    11341 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
    11401 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
    11461 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
    11521 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
    11581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
    11641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
    11701 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
    11761 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
    11821 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
    11881 tgcacacagc ccagcttgga gcgaacgacc t
//
