LOCUS       sequence-273695-       12046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a mouse SIK2-targeting sgRNA, Cas9, and Cre-recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12046)
  AUTHORS   Hollstein PE, Eichner LJ, Brun SN, Kamireddy A, Svensson RU, Vera 
            LI, Ross DS, Rymoff TJ, Hutchins A, Galvez HM, Williams AE, 
            Shokhirev MN, Screaton RA, Berdeaux R, Shaw RJ
  TITLE     The AMPK-Related Kinases SIK1 and SIK3 Mediate Key Tumor-Suppressive
            Effects of LKB1 in NSCLC.
  JOURNAL   Cancer Discov. 2019 Nov;9(11):1606-1627. doi: 
            10.1158/2159-8290.CD-18-1261. Epub 2019 Jul 26.
  PUBMED    31350328
REFERENCE   2  (bases 1 to 12046)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1158/2159-8290.CD-18-1261"; journalName: "Cancer Discov"; date: 
            "2019-11"; volume: "9"; issue: "11"; pages: "1606-1627"
COMMENT     Sequence Label: pSECC-sgSIK2-B
FEATURES             Location/Qualifiers
     source          1..12046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1683..1916
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2101..2145
                     /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    2443..2560
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2660..2908
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2660..2680
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2831..2850
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2929..3004
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3052..3169
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3268..3479
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3524..3533
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3533..3556
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3563..3583
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3608..7708
                     /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             3608..7708
                     /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             7709..7756
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7772..7828
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7832..7852
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7853..8881
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPGIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     misc_feature    8934..9522
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8987..9007)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9405..9416)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9525..9541)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9526..9542)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10051..10231
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(10293..10881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10373..10392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11052..11843,11844..11912))
                     /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     11675..11694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11913..12017)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gggtcgacta gtgatcagtg tgagggagtg
     2641 taaagctggg tttcccagag agggcctatt tcccatgatt ccttcatatt tgcatatacg
     2701 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
     2761 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     2821 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     2881 atatatcttg tggaaaggac gaaacaccgg tggggttcta cgacatcggt tttagagcta
     2941 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     3001 gtgctttttt gaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag
     3061 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     3121 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     3181 cagggacagc agagatccac tttggcgccg gcaaaccgct gttcctagga atcccgaggc
     3241 cttcgagtgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     3301 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
     3361 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     3421 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     3481 tgtcgtgacg cgggacccga cattagcgct acagcttaag cgggccacca tggattacaa
     3541 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
     3601 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     3661 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     3721 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     3781 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     3841 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     3901 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     3961 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     4021 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     4081 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     4141 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     4201 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     4261 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     4321 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     4381 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     4441 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     4501 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     4561 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     4621 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     4681 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     4741 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     4801 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     4861 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     4921 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     4981 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     5041 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     5101 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     5161 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     5221 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     5281 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     5341 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     5401 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     5461 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     5521 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     5581 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     5641 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     5701 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     5761 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     5821 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     5881 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     5941 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     6001 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     6061 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     6121 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     6181 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     6241 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     6301 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     6361 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     6421 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     6481 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     6541 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     6601 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     6661 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     6721 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     6781 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     6841 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     6901 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     6961 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     7021 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     7081 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     7141 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     7201 taagtactcc ctgttcgagc tggaaaacgg caggaagaga atgctggcct ctgccggcga
     7261 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     7321 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     7381 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     7441 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     7501 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     7561 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     7621 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     7681 gatcgacctg tctcagctgg gaggcgacaa gcgtcctgct gctactaaga aagctggtca
     7741 agctaagaaa aagaaagcta gcggcagcgg cgccaccaac ttcagcctgc tgaagcaggc
     7801 cggcgacgtg gaggagaacc ccggccccat gcccaagaag aagaggaagg tgtccaattt
     7861 actgaccgta caccaaaatt tgcctgcatt accggtcgat gcaacgagtg atgaggttcg
     7921 caagaacctg atggacatgt tcagggatcg ccaggcgttt tctgagcata cctggaaaat
     7981 gcttctgtcc gtttgccggt cgtgggcggc atggtgcaag ttgaataacc ggaaatggtt
     8041 tcccgcagaa cctgaagatg ttcgcgatta tcttctatat cttcaggcgc gcggtctggc
     8101 agtaaaaact atccagcaac atttgggcca gctaaacatg cttcatcgtc ggtccgggct
     8161 gccacgacca agtgacagca atgctgtttc actggttatg cggcggatcc gaaaagaaaa
     8221 cgttgatgcc ggtgaacgtg caaaacaggc tctagcgttc gaacgcactg atttcgacca
     8281 ggttcgttca ctcatggaaa atagcgatcg ctgccaggat atacgtaatc tggcatttct
     8341 ggggattgct tataacaccc tgttacgtat agccgaaatt gccaggatca gggttaaaga
     8401 tatctcacgt actgacggtg ggagaatgtt aatccatatt ggcagaacga aaacgctggt
     8461 tagcaccgca ggtgtagaga aggcacttag cctgggggta actaaactgg tcgagcgatg
     8521 gatttccgta tctggtgtag ctgatgatcc gaataactac ctgttttgcc gggtcagaaa
     8581 aaatggtgtt gccgcgccat ctgccaccag ccagctatca actcgcgccc tggaagggat
     8641 ttttgaagca actcatcgat tgatttacgg cgctaaggat gactctggtc agagatacct
     8701 ggcctggtct ggacacagtg cccgtgtcgg agccgcgcga gatatggccc gcgctggagt
     8761 ttcaataccg gggatcatgc aagctggtgg ctggaccaat gtaaatattg tcatgaacta
     8821 tatccgtaac ctggatagtg aaacaggggc aatggtgcgc ctgctggaag atggcgatta
     8881 gaaacgttgt tgtttggggt tgaattactc ttctagaccg cggcgcgccc gataatcaac
     8941 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     9001 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     9061 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     9121 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     9181 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     9241 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     9301 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     9361 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     9421 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     9481 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     9541 gatcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     9601 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     9661 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     9721 aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     9781 atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc
     9841 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta
     9901 gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg
     9961 gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac
    10021 atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc
    10081 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
    10141 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
    10201 cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc
    10261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    10321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    10381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    10441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    10501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    10561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    10621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    10681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    10741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    10801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    10861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    10921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    10981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    11041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    11101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    11161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    11221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    11281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    11341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    11401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    11461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    11521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    11581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    11641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    11701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    11761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    11821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    11881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    11941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    12001 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacg
//
