LOCUS       sequence-374829-       15482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 with an N-term Sid4x fusion, a C-term 
            HA-2xNLS-XTEN80(linker)-KRAB(Kox1) fusion, and GFP linked by a P2A 
            site from a SFFV promoter with an upstream ubiquitous chromatin 
            opening element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15482)
  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 15482)
  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-SFFV-SID4x-dCas9-XTEN80-KRAB(Kox1)-P2...
FEATURES             Location/Qualifiers
     source          1..15482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          72..137
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             267..287
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    712..846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(749..768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     803..822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             1014..1647
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1694..1819
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2316..2549
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2734..2778
                     /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    3063..3180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3922..4329
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             join(4836..4859,4860..4862,4863..7349,7350..7352,
                     7353..8936)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4836..8936
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8940..8966
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8946..8972
                     /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             8985..9005
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9012..9032
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9342..9527
                     /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             9537..9593
                     /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(9594..9596,9597..10310)
                     /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(9897..9916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10244..10265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    10359..10947
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10412..10432)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10830..10841)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10950..10966)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10951..10967)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11057..11290
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(11315..11333)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(11316..11333)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(11623..11642)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(11856..11875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11975..11997
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(12035..12053)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12121..12225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12226..12294,12295..13086)
                     /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(12444..12463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      13257..13845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13746..13765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13999..14016
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(14086..14106)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        14091..14420
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      14271..14406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct acagagattt
       61 aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa
      121 ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg
      181 cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta
      241 ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg
      301 actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg
      361 cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg
      421 aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac atactgtttt
      481 ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa aaattgtgta
      541 cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat agtgccttga
      601 ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
      661 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
      721 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
      781 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
      841 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
      901 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
      961 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg
     1021 gctaattcac tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg
     1081 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
     1141 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
     1201 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     1261 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
     1321 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
     1381 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
     1441 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     1501 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     1561 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     1621 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     1681 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     1741 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     1801 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     1861 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1921 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1981 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     2041 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     2101 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     2161 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     2221 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     2281 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     2341 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     2401 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     2461 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     2521 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     2581 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     2641 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     2701 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     2761 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     2821 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     2881 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     2941 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     3001 agatccattc gattagtgaa cggatctcga cggtatcgcc aaatggcagt attcatccac
     3061 aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     3121 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     3181 cgggtttatt acagggacag cagagatcca gtttggatcg ataagcttga tatcgaattc
     3241 gggaggtggt ccctgcagtt acgccaatga taacccccgc cagaaaaatc ttagtagcct
     3301 tccctttttg ttttccgtgc cccaactcgg cggattgact cggccccttc cggaaacacc
     3361 cgaatcaact tctagtcaaa ttattgttca cgccgcaatg acccacccct ggcccgcgtc
     3421 tgtggaactg acccctggtg tacaggagag ttcgctgctg aaagtggtcc caaaggggta
     3481 ctagttttta agctcccaac tccccctccc ccagcgtctg gaggattcca caccctcgca
     3541 ccgcaggggc gaggaagtgg gcggagtccg gttttggcgc cagccgctga ggctgccaag
     3601 cagaaaagcc accgctgagg agactccggt cactgtcctc gccccgcctc ccccttccct
     3661 ccccttgggg accaccgggc gccacgccgc gaacggtaag tgccgcggtc gtcggcgcct
     3721 ccgccctccc cctagggccc caattcccag cgggcgcggc gcgcggcccc gcgcgcagct
     3781 cccggctccc tcccccttcg gatgtggctt gagctgtagg cgcggagtcc tgcagccccg
     3841 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     3901 aggtttggca agctagctgc agtaacgcca ttttgcaagg catggaaaaa taccaaacca
     3961 agaatagaga agttcagatc aagggcgggt acatgaaaat agctaacgtt gggccaaaca
     4021 ggatatctgc ggtgagcagt ttcggccccg gcccggggcc aagaacagat ggtcaccgca
     4081 gtttcggccc cggcccgagg ccaagaacag atggtcccca gatatggccc aaccctcagc
     4141 agtttcttaa gacccatcag atgtttccag gctcccccaa ggacctgaaa tgaccctgcg
     4201 ccttatttga attaaccaat cagcctgctt ctcgcttctg ttcgcgcgct tctgcttccc
     4261 gagctctata aaagagctca caacccctca ctcggcgcgc cagtcctccg acagactgag
     4321 tcgcccgggg gggatctgga gctctcgaga attctcacgc gtctgcagga tatcaagctt
     4381 gcggtaccgc gcgtacggcc accatgaaca tccaaatgct gctggaagcc gccgactatc
     4441 tggaaaggag ggagagggag gccgaacacg gctacgcctc catgctgccc ggcagcggca
     4501 tgaatatcca aatgctcctg gaggccgctg attacctgga aaggagggaa agagaggctg
     4561 agcacggata tgcctccatg ctccccggct ccggcatgaa tattcagatg ctgctcgagg
     4621 ccgccgacta cctggagagg agggagagag aagccgagca cggctatgcc tccatgctgc
     4681 ctggcagcgg catgaatatc cagatgctgc tggaggccgc cgactacctg gaaagaaggg
     4741 aaagggaggc cgaacatggc tatgccagca tgctgcctgg ttctggcgga gggccgagct
     4801 ctggcgcacc cccaccaagt ggagggtctc ctgccgacaa gaagtacagc atcggcctgg
     4861 ccatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     4921 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggcg
     4981 ccctgctgtt cgacagcgga gaaacagccg aggccacccg gctgaagaga accgccagaa
     5041 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     5101 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     5161 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     5221 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     5281 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     5341 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     5401 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     5461 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     5521 ccggcgagaa gaagaatggc ctgttcggca acctgattgc cctgagcctg ggcctgaccc
     5581 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     5641 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     5701 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     5761 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     5821 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     5881 ttttcttcga ccagagcaag aacggctacg ccggctacat cgatggcgga gccagccagg
     5941 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     6001 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     6061 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     6121 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     6181 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     6241 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgcc agcgcccaga
     6301 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     6361 agcacagcct gctgtacgag tacttcaccg tgtacaacga gctgaccaaa gtgaaatacg
     6421 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaaa gccatcgtgg
     6481 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     6541 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     6601 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     6661 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     6721 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     6781 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     6841 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     6901 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     6961 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     7021 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     7081 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     7141 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     7201 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     7261 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     7321 tcaaccggct gtccgactac gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg
     7381 actccatcga taacaaagtg ctgactcgga gcgacaagaa ccggggcaag agcgacaacg
     7441 tgccctccga agaggtcgtg aagaagatga agaactactg gcgccagctg ctgaatgcca
     7501 agctgattac ccagaggaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     7561 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     7621 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaac gacaaactga
     7681 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     7741 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     7801 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     7861 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     7921 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     7981 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaacag
     8041 gcgagatcgt gtgggataag ggccgggact ttgccaccgt gcggaaagtg ctgtctatgc
     8101 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     8161 tcctgcccaa gaggaacagc gacaagctga tcgccagaaa gaaggactgg gaccctaaga
     8221 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     8281 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     8341 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     8401 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     8461 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     8521 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     8581 aggataatga gcagaaacag ctgtttgtgg aacagcacaa acactacctg gacgagatca
     8641 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaagg
     8701 tgctgagcgc ctacaacaag cacagagaca agcctatcag agagcaggcc gagaatatca
     8761 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     8821 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     8881 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacgcct
     8941 atccctatga cgtgcccgat tatgccagcc tgggcagcgg ctcccccaag aaaaaacgca
     9001 aggtggaaga tcctaagaaa aagcggaaag tggacggcat tggtagtggg agcaacggca
     9061 gcagcggatc cggagggccg agctctggcg cacccccacc aagtggaggg tctcctgccg
     9121 ggtccccaac atctactgaa gaaggcacca gcgaatccgc aacgcccgag tcaggccctg
     9181 gtacctccac agaaccatct gaaggtagtg cgcctggttc cccagctgga agccctactt
     9241 ccaccgaaga aggcacgtca accgaaccaa gtgaaggatc tgcccctggg accagcactg
     9301 aaccatctga gatggatgct aagtcactaa ctgcctggtc ccggacactg gtgaccttca
     9361 aggatgtatt tgtggacttc accagggagg agtggaagct gctggacact gctcagcaga
     9421 tcgtgtacag aaatgtgatg ctggagaact ataagaacct ggtttccttg ggttatcagc
     9481 ttactaagcc agatgtgatc ctccggttgg agaagggaga agagcccagc ggccgcgcta
     9541 ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga cctgtgagca
     9601 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     9661 acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga
     9721 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     9781 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     9841 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     9901 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     9961 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
    10021 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
    10081 cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
    10141 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
    10201 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
    10261 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa taagcgatcg
    10321 ccctgcaggc atgcaagctt gatatcaagc ttatcgataa tcaacctctg gattacaaaa
    10381 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
    10441 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
    10501 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
    10561 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
    10621 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
    10681 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
    10741 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
    10801 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
    10861 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
    10921 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgaggg aattaattcg
    10981 agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
    11041 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
    11101 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
    11161 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
    11221 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
    11281 atctctagca gcatctagaa ttaattccgt gtattctata gtgtcaccta aatcgtatgt
    11341 gtatgataca taaggttatg tattaattgt agccgcgttc taacgacaat atgtacaagc
    11401 ctaattgtgt agcatctggc ttactgaagc agaccctatc atctctctcg taaactgccg
    11461 tcagagtcgg tttggttgga cgaaccttct gagtttctgg taacgccgtc ccgcacccgg
    11521 aaatggtcag cgaaccaatc agcagggtca tcgctagcca gatcctctac gccggacgca
    11581 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
    11641 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
    11701 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
    11761 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
    11821 cgcataaggg agagcgtcga atggtgcact ctcagtacaa tctgctctga tgccgcatag
    11881 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
    11941 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
    12001 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
    12061 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
    12121 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
    12181 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
    12241 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
    12301 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
    12361 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
    12421 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
    12481 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
    12541 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
    12601 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
    12661 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
    12721 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
    12781 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
    12841 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    12901 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    12961 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    13021 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    13081 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    13141 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    13201 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    13261 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
    13321 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
    13381 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
    13441 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
    13501 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
    13561 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    13621 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    13681 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    13741 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    13801 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    13861 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
    13921 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
    13981 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
    14041 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgtggaat gtgtgtcagt
    14101 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
    14161 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
    14221 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
    14281 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
    14341 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
    14401 gaggcctagg cttttgcaaa aagcttggac acaagacagg cttgcgagat atgtttgaga
    14461 ataccacttt atcccgcgtc agggagaggc agtgcgtaaa aagacgcgga ctcatgtgaa
    14521 atactggttt ttagtgcgcc agatctctat aatctcgcgc aacctatttt cccctcgaac
    14581 actttttaag ccgtagataa acaggctggg acacttcaca tgagcgaaaa atacatcgtc
    14641 acctgggaca tgttgcagat ccatgcacgt aaactcgcaa gccgactgat gccttctgaa
    14701 caatggaaag gcattattgc cgtaagccgt ggcggtctgt accgggtgcg ttactggcgc
    14761 gtgaactggg tattcgtcat gtcgataccg tttgtatttc cagctacgat cacgacaacc
    14821 agcgcgagct taaagtgctg aaacgcgcag aaggcgatgg cgaaggcttc atcgttattg
    14881 atgacctggt ggataccggt ggtactgcgg ttgcgattcg tgaaatgtat ccaaaagcgc
    14941 actttgtcac catcttcgca aaaccggctg gtcgtccgct ggttgatgac tatgttgttg
    15001 atatcccgca agatacctgg attgaacagc cgtgggatat gggcgtcgta ttcgtcccgc
    15061 caatctccgg tcgctaatct tttcaacgcc tggcactgcc gggcgttgtt ctttttaact
    15121 tcaggcgggt tacaatagtt tccagtaagt attctggagg ctgcatccat gacacaggca
    15181 aacctgagcg aaaccctgtt caaaccccgc tttaaacatc ctgaaacctc gacgctagtc
    15241 cgccgcttta atcacggcgc acaaccgcct gtgcagtcgg cccttgatgg taaaaccatc
    15301 cctcactggt atcgcatgat taaccgtctg atgtggatct ggcgcggcat tgacccacgc
    15361 gaaatcctcg acgtccaggc acgtattgtg atgagcgatg ccgaacgtac cgacgatgat
    15421 ttatacgata cggtgattgg ctaccgtggc ggcaactgga tttatgagtg ggccccggat
    15481 ct
//
