LOCUS       sequence-205822-       13102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 engineered chromatin immunoprecipitation of D4Z4 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13102)
  AUTHORS   Campbell AE, Shadle SC, Jagannathan S, Lim JW, Resnick R, Tawil R, 
            van der Maarel SM, Tapscott SJ
  TITLE     NuRD and CAF-1-mediated silencing of the D4Z4 array is modulated by 
            DUX4-induced MBD3L proteins.
  JOURNAL   Elife. 2018 Mar 13;7. pii: 31023. doi: 10.7554/eLife.31023.
  PUBMED    29533181
REFERENCE   2  (bases 1 to 13102)
  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: "2018-03-13"; volume: "7. pii: 31023. doi"; pages: " 
            10.7554/eLife.31023"
COMMENT     Sequence Label: pZLCv2-gD4Z4-3-3xFLAG-dCas9-HA-2xNLS
FEATURES             Location/Qualifiers
     source          1..13102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2371..2619
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2371..2391
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2542..2561
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2641..2716
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2778..2989
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3010..3075
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(3082..3105,3106..3108,3109..5595,5596..5598,
                     5599..7182)
                     /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             3082..7182
                     /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             7186..7212
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7192..7218
                     /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             7231..7251
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7258..7278
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7288..7344
                     /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             7345..7941
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7838..7858
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7957..8545
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8010..8030)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8428..8439)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8617..8850
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8876..8893)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8882..9106
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9152..9580
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9239..9258)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9449..9470
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9589..9609)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9594..9923
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9774..9909
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9836..9855
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9971..10018
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10037..10411
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10541..10662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10578..10597)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10632..10651
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10711..10727)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10711..10727)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10724..10746)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10735..10751
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(10759..10765,10766..10783,10784..10789))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10804..10825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10942..10959)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11113..11701)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11193..11212)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11872..12663,12664..12732))
                     /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     12495..12514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12733..12837)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12912..12931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
     2401 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2461 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2521 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2581 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gtgagcaaca ggccgccttg
     2641 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     2701 ggcaccgagt cggtgctttt ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg
     2761 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     2821 aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     2881 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     2941 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg accggtgccg
     3001 cccaccatgg actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag
     3061 gatgacgatg acaaggaatt cgacaagaag tacagcatcg gcctggccat cggcaccaac
     3121 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     3181 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggcgccct gctgttcgac
     3241 agcggagaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     3301 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     3361 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     3421 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     3481 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     3541 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     3601 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     3661 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     3721 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     3781 aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     3841 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     3901 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     3961 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     4021 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     4081 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     4141 agcaagaacg gctacgccgg ctacatcgat ggcggagcca gccaggaaga gttctacaag
     4201 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     4261 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     4321 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     4381 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     4441 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     4501 ccctggaact tcgaggaagt ggtggacaag ggcgccagcg cccagagctt catcgagcgg
     4561 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     4621 tacgagtact tcaccgtgta caacgagctg accaaagtga aatacgtgac cgagggaatg
     4681 agaaagcccg ccttcctgag cggcgagcag aaaaaagcca tcgtggacct gctgttcaag
     4741 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     4801 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     4861 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     4921 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     4981 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     5041 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     5101 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     5161 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     5221 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     5281 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     5341 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     5401 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     5461 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     5521 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     5581 gactacgatg tggacgctat cgtgcctcag agctttctga aggacgactc catcgataac
     5641 aaagtgctga ctcggagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     5701 gtcgtgaaga agatgaagaa ctactggcgc cagctgctga atgccaagct gattacccag
     5761 aggaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     5821 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     5881 ctggactccc ggatgaacac taagtacgac gagaacgaca aactgatccg ggaagtgaaa
     5941 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     6001 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     6061 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     6121 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     6181 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     6241 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaacaggcga gatcgtgtgg
     6301 gataagggcc gggactttgc caccgtgcgg aaagtgctgt ctatgcccca agtgaatatc
     6361 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     6421 aacagcgaca agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     6481 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     6541 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     6601 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     6661 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     6721 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     6781 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     6841 aaacagctgt ttgtggaaca gcacaaacac tacctggacg agatcatcga gcagatcagc
     6901 gagttctcca agagagtgat cctggccgac gctaatctgg acaaggtgct gagcgcctac
     6961 aacaagcaca gagacaagcc tatcagagag caggccgaga atatcatcca cctgtttacc
     7021 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     7081 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     7141 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acgcctatcc ctatgacgtg
     7201 cccgattatg ccagcctggg cagcggctcc cccaagaaaa aacgcaaggt ggaagatcct
     7261 aagaaaaagc ggaaagtggg atccggcgca acaaacttct ctctgctgaa acaagccgga
     7321 gatgtcgaag agaatcctgg accgaccgag tacaagccca cggtgcgcct cgccacccgc
     7381 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     7441 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     7501 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     7561 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     7621 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     7681 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac
     7741 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     7801 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     7861 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     7921 acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt
     7981 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     8041 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     8101 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     8161 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     8221 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     8281 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     8341 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     8401 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     8461 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     8521 atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg
     8581 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     8641 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     8701 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     8761 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     8821 ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg
     8881 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     8941 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     9001 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     9061 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     9121 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     9181 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     9241 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     9301 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     9361 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     9421 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     9481 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     9541 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     9601 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     9661 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     9721 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     9781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     9841 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     9901 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     9961 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
    10021 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
    10081 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
    10141 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
    10201 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
    10261 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
    10321 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
    10381 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
    10441 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
    10501 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
    10561 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
    10621 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
    10681 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
    10741 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
    10801 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
    10861 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
    10921 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
    10981 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
    11041 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
    11101 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
    11161 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
    11221 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
    11281 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
    11341 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
    11401 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
    11461 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
    11521 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
    11581 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
    11641 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
    11701 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    11761 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    11821 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    11881 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    11941 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    12001 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    12061 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    12121 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    12181 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    12241 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    12301 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    12361 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    12421 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    12481 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    12541 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    12601 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    12661 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    12721 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    12781 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    12841 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
    12901 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
    12961 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
    13021 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
    13081 gtacgggcca gatatacgcg tt
//
