LOCUS       sequence-366451-       11911 bp    DNA     circular SYN 15-APR-2025
DEFINITION  gRNA targeting mouse cGAS (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 Apr 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 cGAS #2-gRNA-Cas9-Puro
FEATURES             Location/Qualifiers
     source          1..11911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     277..297
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    393..981
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(446..466)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(864..875)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1068..1301
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1373..1494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1410..1429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1464..1483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1534..1669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1689..1708)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1690..1708)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1718..1735)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1718..1734)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1727..1749)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1876..2331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1963..1982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2173..2194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2357..2461
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2462..2530,2531..3322)
                     /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(2680..2699)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3493..4081
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3982..4001
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4235..4252
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4369..4390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4405..4410,4411..4428,4429..4435)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4443..4459
                     /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     4448..4470
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4467..4483
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4467..4483
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4502..4522
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4504..4522
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4550..4776
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4777..4957
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5004..5129
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5622..5855
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6040..6084
                     /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    6351..6468
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6542..6790
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6542..6562
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6713..6732
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6811..6886
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(6898..6922)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     7263..7287
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      7357..7366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7366..7431
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7438..7458
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7483..11583
                     /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             7483..11583
                     /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             11584..11631
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             11641..11694
                     /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             join(11695..11911,1..380)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
ORIGIN
        1 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
       61 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
      121 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
      181 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
      241 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
      301 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
      361 cccgcaagcc cggtgcctga actagtgtcg acaatcaacc tctggattac aaaatttgtg
      421 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
      481 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
      541 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
      601 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
      661 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
      721 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
      781 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
      841 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
      901 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
      961 ccctttgggc cgcctccccg cctggaattc gagctcggta cctttaagac caatgactta
     1021 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     1081 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
     1141 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     1201 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     1261 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     1321 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     1381 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1441 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1501 ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     1561 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     1621 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac
     1681 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     1741 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1801 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1861 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1921 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1981 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2041 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2101 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2161 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2221 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2281 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     2341 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2401 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2461 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2521 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2581 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2641 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2701 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2761 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2821 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2881 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2941 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3001 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3061 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3121 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3181 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3241 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3301 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3361 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3421 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3481 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3541 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3601 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3661 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3721 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3781 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3841 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3901 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3961 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4021 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4081 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4141 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4201 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4261 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     4321 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     4381 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     4441 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
     4501 cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat
     4561 gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga
     4621 gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta
     4681 ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga
     4741 gatattgtat ttaagtgcct aactcgatac ataaacgggt ctctctggtt agaccagatc
     4801 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     4861 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     4921 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     4981 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     5041 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     5101 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     5161 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     5221 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     5281 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     5341 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     5401 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     5461 agcaagcgga cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     5521 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     5581 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     5641 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     5701 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     5761 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     5821 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     5881 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     5941 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     6001 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     6061 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     6121 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     6181 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     6241 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     6301 agagacagat ccattcgatt agtgaacgga tctcgacggt atcggttaac ttttaaaaga
     6361 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     6421 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttat cgtgtacaaa
     6481 aaagcaggct ttaaaggaac caattcagtc gactggatcc ggtaccaagg tcgggcagga
     6541 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     6601 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     6661 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     6721 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     6781 acgaaacacc ggcagcccag agcgccgcga gttttagagc tagaaatagc aagttaaaat
     6841 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttctagacc
     6901 cagctttctt gtacaaagtt ggcattacga ttacgcgtag gaacagagaa acaggagaat
     6961 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     7021 ttggaacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     7081 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     7141 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     7201 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct ctatataagc
     7261 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacttc
     7321 catagaagga tctcgagcgg ccgcgctagc ggatccgcca ccatggacta taaggaccac
     7381 gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca
     7441 aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc
     7501 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     7561 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     7621 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     7681 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     7741 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     7801 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     7861 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     7921 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     7981 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     8041 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     8101 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     8161 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     8221 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     8281 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     8341 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     8401 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     8461 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     8521 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     8581 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     8641 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     8701 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     8761 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     8821 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     8881 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     8941 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     9001 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     9061 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     9121 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     9181 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     9241 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     9301 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     9361 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     9421 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     9481 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     9541 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     9601 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     9661 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     9721 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     9781 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     9841 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     9901 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     9961 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
    10021 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
    10081 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
    10141 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
    10201 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
    10261 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
    10321 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
    10381 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
    10441 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
    10501 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
    10561 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
    10621 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
    10681 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
    10741 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
    10801 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
    10861 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
    10921 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
    10981 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
    11041 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
    11101 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
    11161 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
    11221 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
    11281 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
    11341 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
    11401 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
    11461 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
    11521 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
    11581 gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gggatccggt
    11641 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatccggg ccctaccgag
    11701 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
    11761 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
    11821 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
    11881 gtgtgggtcg cggacgacgg cgccgcggtg g
//
