LOCUS       sequence-343565-       14080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9 repressor KRAB-dCas9-MeCP2 and blasticidin 
            resistance .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14080)
  AUTHORS   Morris JA, Caragine C, Daniloski Z, Domingo J, Barry T, Lu L, Davis 
            K, Ziosi M, Glinos DA, Hao S, Mimitou EP, Smibert P, Roeder K, 
            Katsevich E, Lappalainen T, Sanjana NE
  TITLE     Discovery of target genes and pathways at GWAS loci by pooled 
            single-cell CRISPR screens.
  JOURNAL   Science. 2023 May 19;380(6646):eadh7699. doi: 
            10.1126/science.adh7699. Epub 2023 May 19.
  PUBMED    37141313
REFERENCE   2  (bases 1 to 14080)
  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: "Science."; 
            date: "2023-05-19"; volume: "380(6646):eadh7699. doi"; pages: " 
            10.1126/science.adh7699. Epub 2023 May 19"
COMMENT     Sequence Label: lentiCRISPRi(v2)-Blast
FEATURES             Location/Qualifiers
     source          1..14080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..71)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        243..622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        624..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     777..797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             840..1020
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1067..1192
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2103..2147
                     /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    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2655..2866
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2885..2894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2921..3106
                     /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             join(3128..3154,3155..3157,3158..5644,5645..5647,
                     5648..7231)
                     /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             3128..7231
                     /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             7235..7261
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7241..7267
                     /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             7280..7300
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7307..7327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7400..7420
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7430..8287
                     /codon_start=1
                     /product="transcriptional repression domain from rat 
                     methyl-CpG-binding protein 2 (Nan et al., 1997)"
                     /label=MeCP2
                     /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP
                     GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE
                     TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH
                     HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP
                     KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV
                     TERVS"
     CDS             8297..8353
                     /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             8354..8752
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8777..9365
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8830..8850)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9248..9259)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9368..9384)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9369..9385)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9890..10070
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10096..10113)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10102..10326
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10372..10800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10459..10478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10669..10690
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10809..10829)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10814..11143
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10994..11129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11056..11075
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11191..11238
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11257..11631
                     /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    11761..11882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11798..11817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11852..11871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11931..11947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11931..11947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11944..11966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11955..11971
                     /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(11979..11985,11986..12003,12004..12009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12024..12045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12162..12179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12333..12921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12413..12432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13092..13883,13884..13952))
                     /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     13715..13734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13953..14057)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
       61 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      121 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
      181 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      241 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      301 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      361 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      421 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      481 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      541 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      601 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      661 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      721 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      781 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg
      841 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
      901 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
      961 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     1021 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     1081 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     1141 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     1201 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     1261 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     1321 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     1381 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     1441 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     1501 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt aattagctag ctaggtcttg aaaggagtgg gaattggctc
     2641 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     2701 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     2761 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     2821 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacc ggttctagag
     2881 cgctgccacc atggatgcta agtcactaac tgcctggtcc cggacactgg tgaccttcaa
     2941 ggatgtattt gtggacttca ccagggagga gtggaagctg ctggacactg ctcagcagat
     3001 cgtgtacaga aatgtgatgc tggagaacta taagaacctg gtttccttgg gttatcagct
     3061 tactaagcca gatgtgatcc tccggttgga gaagggagaa gagcccggcg gttccggcgg
     3121 agggtcgatg gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
     3181 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     3241 cgaccggcac agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggagaaac
     3301 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     3361 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     3421 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     3481 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     3541 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     3601 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     3661 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     3721 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     3781 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     3841 cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     3901 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     3961 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     4021 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     4081 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     4141 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     4201 ctacgccggc tacatcgatg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     4261 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     4321 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     4381 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     4441 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     4501 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     4561 cgaggaagtg gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt
     4621 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     4681 caccgtgtac aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     4741 cttcctgagc ggcgagcaga aaaaagccat cgtggacctg ctgttcaaga ccaaccggaa
     4801 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     4861 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     4921 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     4981 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     5041 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     5101 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     5161 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     5221 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     5281 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     5341 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     5401 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     5461 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     5521 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     5581 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     5641 ggacgctatc gtgcctcaga gctttctgaa ggacgactcc atcgataaca aagtgctgac
     5701 tcggagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     5761 gatgaagaac tactggcgcc agctgctgaa tgccaagctg attacccaga ggaagttcga
     5821 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     5881 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     5941 gatgaacact aagtacgacg agaacgacaa actgatccgg gaagtgaaag tgatcaccct
     6001 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     6061 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     6121 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     6181 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     6241 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     6301 gaagcggcct ctgatcgaga caaacggcga aacaggcgag atcgtgtggg ataagggccg
     6361 ggactttgcc accgtgcgga aagtgctgtc tatgccccaa gtgaatatcg tgaaaaagac
     6421 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgacaa
     6481 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     6541 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     6601 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     6661 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     6721 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     6781 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     6841 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     6901 tgtggaacag cacaaacact acctggacga gatcatcgag cagatcagcg agttctccaa
     6961 gagagtgatc ctggccgacg ctaatctgga caaggtgctg agcgcctaca acaagcacag
     7021 agacaagcct atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     7081 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     7141 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     7201 acggatcgac ctgtctcagc tgggaggcga cgcctatccc tatgacgtgc ccgattatgc
     7261 cagcctgggc agcggctccc ccaagaaaaa acgcaaggtg gaagatccta agaaaaagcg
     7321 gaaagtggac ggcattggta gtgggagcaa cggcagcagc gggtcctcgg gaggtggttc
     7381 gggtggctct ggatcaagcc caaagaagaa acggaaggtg gaagcctcag tgcaggtgaa
     7441 aagggtgctg gaaaaatccc ccggcaaact cctcgtgaag atgcccttcc aggcttcccc
     7501 tggcggaaaa ggtgaagggg gtggcgcaac cacatctgcc caggtcatgg tcatcaagcg
     7561 acctggaagg aaaagaaagg ccgaggctga ccctcaggcc attccaaaga aacggggacg
     7621 caagccaggg tccgtggtcg cagctgcagc agctgaggct aagaaaaagg cagtgaagga
     7681 aagctccatc cgcagtgtgc aggagactgt cctgcccatc aagaagagga agactaggga
     7741 gaccgtgtcc atcgaggtca aagaagtggt caagcccctg ctcgtgtcca ccctgggcga
     7801 aaaatctgga aaggggctca aaacatgcaa gtcacctgga cggaaaagca aggagtctag
     7861 tccaaagggg cgctcaagct ccgcttctag tccccctaaa aaggaacacc atcaccatca
     7921 ccatcacgcc gagtctccta aggctcctat gccactgctc ccaccacctc caccacctga
     7981 gccacagtca agcgaagacc ccatcagccc acccgagcct caggatctgt cctctagtat
     8041 ttgcaaagag gaaaagatgc ccagagcagg cagcctggag agtgatggct gtccaaaaga
     8101 acccgccaag acccagccta tggtggcagc cgctgcaact accaccacaa ccacaactac
     8161 cacagtggcc gaaaaataca agcatcgcgg cgagggcgaa cgaaaggaca ttgtgtcaag
     8221 ctccatgccc agacctaacc gggaggaacc agtcgatagt aggacacccg tgactgagag
     8281 agtctcagga tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga
     8341 gaatcctgga ccgatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     8401 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     8461 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     8521 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     8581 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     8641 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     8701 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagaattcga
     8761 tatcaagctt atcggtaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     8821 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     8881 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     8941 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     9001 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     9061 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     9121 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     9181 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     9241 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     9301 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     9361 cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca caagtagcaa
     9421 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     9481 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     9541 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     9601 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
     9661 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
     9721 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
     9781 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
     9841 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
     9901 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     9961 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    10021 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
    10081 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
    10141 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
    10201 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
    10261 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
    10321 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
    10381 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
    10441 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    10501 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
    10561 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
    10621 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    10681 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
    10741 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
    10801 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
    10861 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
    10921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
    10981 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
    11041 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
    11101 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
    11161 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
    11221 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
    11281 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
    11341 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
    11401 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
    11461 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
    11521 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
    11581 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
    11641 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
    11701 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
    11761 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    11821 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    11881 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
    11941 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
    12001 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
    12061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
    12121 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
    12181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
    12241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    12301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    12361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    12421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    12481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    12541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    12601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    12661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    12721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    12781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    12841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    12901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    12961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    13021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    13081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    13141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    13201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    13261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    13321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    13381 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    13441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    13501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    13561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    13621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    13681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    13741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    13801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    13861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    13921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    13981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    14041 caaatagggg ttccgcgcac atttccccga aaagtgccac
//
