LOCUS       sequence-374872-       15165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 with a C-term HA-2xNLS-XTEN80(linker)-KRAB(Kox1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15165)
  AUTHORS   Replogle JM, Bonnar JL, Pogson AN, Liem CR, Maier NK, Ding Y, 
            Russell BJ, Wang X, Leng K, Guna A, Norman TM, Pak RA, Ramos DM, 
            Ward ME, Gilbert LA, Kampmann M, Weissman JS, Jost M
  TITLE     Maximizing CRISPRi efficacy and accessibility with dual-sgRNA 
            libraries and optimal effectors.
  JOURNAL   eLife. 2022 Dec 28;11:e81856. doi: 10.7554/eLife.81856.
  PUBMED    36576240
REFERENCE   2  (bases 1 to 15165)
  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: "eLife."; 
            date: "2022-12-28"; volume: "11:e81856. doi"; pages: " 
            10.7554/eLife.81856"
COMMENT     Sequence Label: pHR-UCOE-CMV-dCas9-XTEN80-KRAB(Kox1)-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..15165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(42..60)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(43..60)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(350..369)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(583..602)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     702..724
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(762..780)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        848..952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(953..1021,1022..1813)
                     /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(1171..1190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1984..2572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2473..2492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2726..2743
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2813..2833)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2818..3147
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2998..3133
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4281..4346
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4476..4496
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4921..5055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4958..4977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5012..5031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5223..5856
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5903..6028
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6525..6758
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6943..6987
                     /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    7272..7389
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        8047..8425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8426..8629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8580..8600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8626..8650
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      8719..8728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(8725..8751,8752..8754,8755..11241,11242..11244,
                     11245..12828)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8725..12828
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             12832..12858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             12838..12864
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             12877..12897
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12904..12924
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13234..13419
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             13429..13485
                     /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             join(13486..13488,13489..14202)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(13789..13808)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     14136..14157
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    14251..14839
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(14304..14324)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14722..14733)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(14842..14858)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14843..14859)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             join(14949..15165,1..17)
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 gtggaaaatc tctagcagca tctagaatta attccgtgta ttctatagtg tcacctaaat
       61 cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg
      121 tacaagccta attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa
      181 actgccgtca gagtcggttt ggttggacga accttctgag tttctggtaa cgccgtcccg
      241 cacccggaaa tggtcagcga accaatcagc agggtcatcg ctagccagat cctctacgcc
      301 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
      361 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
      421 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
      481 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
      541 caggagtcgc ataagggaga gcgtcgaatg gtgcactctc agtacaatct gctctgatgc
      601 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      661 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
      721 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
      781 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
      841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      961 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1021 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1081 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1141 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1201 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1261 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1321 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1381 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1441 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1501 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1561 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1621 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1681 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1741 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1801 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1861 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1921 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2101 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2161 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2221 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2281 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2341 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2401 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2461 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2521 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2581 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2641 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2701 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2761 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct gtggaatgtg
     2821 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2881 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     2941 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3001 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3061 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3121 ttttttggag gcctaggctt ttgcaaaaag cttggacaca agacaggctt gcgagatatg
     3181 tttgagaata ccactttatc ccgcgtcagg gagaggcagt gcgtaaaaag acgcggactc
     3241 atgtgaaata ctggttttta gtgcgccaga tctctataat ctcgcgcaac ctattttccc
     3301 ctcgaacact ttttaagccg tagataaaca ggctgggaca cttcacatga gcgaaaaata
     3361 catcgtcacc tgggacatgt tgcagatcca tgcacgtaaa ctcgcaagcc gactgatgcc
     3421 ttctgaacaa tggaaaggca ttattgccgt aagccgtggc ggtctgtacc gggtgcgtta
     3481 ctggcgcgtg aactgggtat tcgtcatgtc gataccgttt gtatttccag ctacgatcac
     3541 gacaaccagc gcgagcttaa agtgctgaaa cgcgcagaag gcgatggcga aggcttcatc
     3601 gttattgatg acctggtgga taccggtggt actgcggttg cgattcgtga aatgtatcca
     3661 aaagcgcact ttgtcaccat cttcgcaaaa ccggctggtc gtccgctggt tgatgactat
     3721 gttgttgata tcccgcaaga tacctggatt gaacagccgt gggatatggg cgtcgtattc
     3781 gtcccgccaa tctccggtcg ctaatctttt caacgcctgg cactgccggg cgttgttctt
     3841 tttaacttca ggcgggttac aatagtttcc agtaagtatt ctggaggctg catccatgac
     3901 acaggcaaac ctgagcgaaa ccctgttcaa accccgcttt aaacatcctg aaacctcgac
     3961 gctagtccgc cgctttaatc acggcgcaca accgcctgtg cagtcggccc ttgatggtaa
     4021 aaccatccct cactggtatc gcatgattaa ccgtctgatg tggatctggc gcggcattga
     4081 cccacgcgaa atcctcgacg tccaggcacg tattgtgatg agcgatgccg aacgtaccga
     4141 cgatgattta tacgatacgg tgattggcta ccgtggcggc aactggattt atgagtgggc
     4201 cccggatctt tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta
     4261 cagagattta aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac
     4321 tgattctaat tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt
     4381 ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg
     4441 atgaggctac tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag
     4501 accccaagga ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata
     4561 gaactcttgc ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga
     4621 aaattatgga aaaatattct gtaaccttta taagtaggca taacagttat aatcataaca
     4681 tactgttttt tcttactcca cacaggcata gagtgtctgc tattaataac tatgctcaaa
     4741 aattgtgtac ctttagcttt ttaatttgta aaggggttaa taaggaatat ttgatgtata
     4801 gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     4861 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4921 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4981 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5041 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
     5101 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
     5161 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
     5221 gttggaaggg ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca
     5281 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     5341 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     5401 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     5461 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     5521 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     5581 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     5641 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     5701 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5761 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5821 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     5881 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     5941 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     6001 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6061 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6121 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6181 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6241 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6301 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     6361 gcaagcggcc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     6421 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     6481 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     6541 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     6601 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     6661 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     6721 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     6781 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     6841 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     6901 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     6961 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     7021 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     7081 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     7141 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     7201 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcca aatggcagta
     7261 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     7321 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     7381 caaaattttc gggtttatta cagggacagc agagatccag tttggatcga taagcttgat
     7441 atcgaattcg ggaggtggtc cctgcagtta cgccaatgat aacccccgcc agaaaaatct
     7501 tagtagcctt ccctttttgt tttccgtgcc ccaactcggc ggattgactc ggccccttcc
     7561 ggaaacaccc gaatcaactt ctagtcaaat tattgttcac gccgcaatga cccacccctg
     7621 gcccgcgtct gtggaactga cccctggtgt acaggagagt tcgctgctga aagtggtccc
     7681 aaaggggtac tagtttttaa gctcccaact ccccctcccc cagcgtctgg aggattccac
     7741 accctcgcac cgcaggggcg aggaagtggg cggagtccgg ttttggcgcc agccgctgag
     7801 gctgccaagc agaaaagcca ccgctgagga gactccggtc actgtcctcg ccccgcctcc
     7861 cccttccctc cccttgggga ccaccgggcg ccacgccgcg aacggtaagt gccgcggtcg
     7921 tcggcgcctc cgccctcccc ctagggcccc aattcccagc gggcgcggcg cgcggccccg
     7981 cgcgcagctc ccggctccct cccccttcgg atgtggcttg agctgtaggc gcggagtcct
     8041 gcagccacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     8101 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     8161 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     8221 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     8281 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     8341 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     8401 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     8461 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     8521 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     8581 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     8641 ccgtcagatc ggggatctgg agctctcgag aattctcacg cgtctgcagg atatcaagct
     8701 tgcggtaccg cgggcccggc caccatggac aagaagtaca gcatcggcct ggccatcggc
     8761 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
     8821 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg cgccctgctg
     8881 ttcgacagcg gagaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
     8941 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
     9001 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
     9061 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
     9121 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
     9181 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     9241 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     9301 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     9361 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     9421 aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac ccccaacttc
     9481 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     9541 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     9601 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     9661 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     9721 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     9781 gaccagagca agaacggcta cgccggctac atcgatggcg gagccagcca ggaagagttc
     9841 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     9901 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     9961 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
    10021 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
    10081 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
    10141 atcaccccct ggaacttcga ggaagtggtg gacaagggcg ccagcgccca gagcttcatc
    10201 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
    10261 ctgctgtacg agtacttcac cgtgtacaac gagctgacca aagtgaaata cgtgaccgag
    10321 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aagccatcgt ggacctgctg
    10381 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
    10441 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
    10501 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
    10561 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
    10621 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
    10681 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
    10741 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
    10801 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
    10861 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
    10921 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
    10981 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
    11041 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
    11101 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
    11161 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
    11221 ctgtccgact acgatgtgga cgctatcgtg cctcagagct ttctgaagga cgactccatc
    11281 gataacaaag tgctgactcg gagcgacaag aaccggggca agagcgacaa cgtgccctcc
    11341 gaagaggtcg tgaagaagat gaagaactac tggcgccagc tgctgaatgc caagctgatt
    11401 acccagagga agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
    11461 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
    11521 cagatcctgg actcccggat gaacactaag tacgacgaga acgacaaact gatccgggaa
    11581 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
    11641 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
    11701 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
    11761 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
    11821 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
    11881 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac aggcgagatc
    11941 gtgtgggata agggccggga ctttgccacc gtgcggaaag tgctgtctat gccccaagtg
    12001 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
    12061 aagaggaaca gcgacaagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
    12121 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
    12181 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
    12241 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
    12301 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
    12361 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
    12421 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
    12481 gagcagaaac agctgtttgt ggaacagcac aaacactacc tggacgagat catcgagcag
    12541 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa ggtgctgagc
    12601 gcctacaaca agcacagaga caagcctatc agagagcagg ccgagaatat catccacctg
    12661 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
    12721 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
    12781 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacgc ctatccctat
    12841 gacgtgcccg attatgccag cctgggcagc ggctccccca agaaaaaacg caaggtggaa
    12901 gatcctaaga aaaagcggaa agtggacggc attggtagtg ggagcaacgg cagcagcgga
    12961 tccggagggc cgagctctgg cgcaccccca ccaagtggag ggtctcctgc cgggtcccca
    13021 acatctactg aagaaggcac cagcgaatcc gcaacgcccg agtcaggccc tggtacctcc
    13081 acagaaccat ctgaaggtag tgcgcctggt tccccagctg gaagccctac ttccaccgaa
    13141 gaaggcacgt caaccgaacc aagtgaagga tctgcccctg ggaccagcac tgaaccatct
    13201 gagatggatg ctaagtcact aactgcctgg tcccggacac tggtgacctt caaggatgta
    13261 tttgtggact tcaccaggga ggagtggaag ctgctggaca ctgctcagca gatcgtgtac
    13321 agaaatgtga tgctggagaa ctataagaac ctggtttcct tgggttatca gcttactaag
    13381 ccagatgtga tcctccggtt ggagaaggga gaagagccca gcggccgcgc tactaacttc
    13441 agcctgctga agcaggctgg agacgtggag gagaaccctg gacctgtgag caagggcgag
    13501 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
    13561 aagttcagcg tgtctggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
    13621 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
    13681 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
    13741 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
    13801 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
    13861 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
    13921 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggcgaacttc
    13981 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
    14041 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
    14101 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
    14161 gccgccggga tcactctcgg catggacgag ctgtacaagt aataagcgat cgccctgcag
    14221 gcatgcaagc ttgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
    14281 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
    14341 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
    14401 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
    14461 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
    14521 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
    14581 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
    14641 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
    14701 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
    14761 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
    14821 ctttgggccg cctccccgca tcgataccgt cgacctcgag ggaattaatt cgagctcggt
    14881 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
    14941 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
    15001 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
    15061 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
    15121 gtgtgactct ggtaactaga gatccctcag acccttttag tcagt
//
