LOCUS       sequence-438981-       16409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2nd Generation Lentiviral vector. Expresses an N-terminal LSD1-dCas9
            fusion protein and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16409)
  AUTHORS   Li K, Liu Y, Cao H, Zhang Y, Gu Z, Liu X, Yu A, Kaphle P, Dickerson 
            KE, Ni M, Xu J
  TITLE     Interrogation of enhancer function by enhancer-targeting CRISPR 
            epigenetic editing.
  JOURNAL   Nat Commun. 2020 Jan 24;11(1):485. doi: 10.1038/s41467-020-14362-5.
  PUBMED    31980609
REFERENCE   2  (bases 1 to 16409)
  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: "Nat 
            Commun."; date: "2020-01-24"; volume: "11(1):485. doi"; pages: " 
            10.1038/s41467-020-14362-5"
COMMENT     Sequence Label: pHR_TRE3G-LSD1-dCas9-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..16409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2050..2167
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2229..2578
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    2237..2255
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2273..2291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2309..2327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2345..2363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2381..2399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2417..2435
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2453..2471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2517..2541
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(5209..5235,5236..5238,5239..7725,7726..7728,
                     7729..9312)
                     /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             5209..9312
                     /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             9316..9342
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9322..9348
                     /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             9361..9381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9388..9408
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9448..9504
                     /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             9505..10215
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(9650..9668)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9918..9937
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    10273..10861
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10326..10346)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10744..10755)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10864..10880)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10865..10881)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10971..11204
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(11229..11247)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(11230..11247)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(11537..11556)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(11770..11789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11889..11911
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(11949..11967)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12035..12139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12140..12208,12209..13000)
                     /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(12358..12377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      13171..13759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13660..13679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13913..13930
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(14000..14020)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        14005..14334
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      14185..14320
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          15468..15533
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             15663..15683
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    16108..16242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(16145..16164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     16199..16218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccaaa tggcagtatt
     2041 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2101 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2161 aaattttcgg gtttattaca gggacagcag agatccagtt tatcgatgag gccctttcgt
     2221 cttcactcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
     2281 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
     2341 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
     2401 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
     2461 agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gcgcctataa aagcagagct
     2521 cgtttagtga accgtcagat cgcctggagc aattccacaa cacttttgtc ttatacttac
     2581 gcgtctgcag gatatcaagc ttgcggtacc gcgggcccgg gatccgccac catgttatct
     2641 gggaagaagg cggcagccgc ggcggcggcg gctgcagcgg cagcaaccgg gacggaggct
     2701 ggccctggga cagcaggcgg ctccgagaac gggtctgagg tggccgcgca gcccgcgggc
     2761 ctgtcgggcc cagccgaggt cgggccgggg gcggtggggg agcgcacacc ccgcaagaaa
     2821 gagcctccgc gggcctcgcc ccccgggggc ctggcggaac cgccggggtc cgcagggcct
     2881 caggccggcc ctactgtcgt gcctgggtct gcgaccccca tggaaactgg aatagcagag
     2941 actccggagg ggcgtcggac cagccggcgc aagcgggcga aggtagagta cagagagatg
     3001 gatgaaagct tggccaacct ctcagaagat gagtattatt cagaagaaga gagaaatgcc
     3061 aaagcagaga aggaaaagaa gcttccccca ccaccccctc aagccccacc tgaggaagaa
     3121 aatgaaagtg agcctgaaga accatcgggt gtggagggcg cagctttcca gagccgactt
     3181 cctcatgacc ggatgacttc tcaagaagca gcctgttttc cagatattat cagtggacca
     3241 caacagaccc agaaggtttt tcttttcatt agaaaccgca cactgcagtt gtggttggat
     3301 aatccaaaga ttcagctgac atttgaggct actctccaac aattagaagc accttataac
     3361 agtgatactg tgcttgtcca ccgagttcac agttatttag agcgtcatgg tcttatcaac
     3421 ttcggcatct ataagaggat aaaaccccta ccaactaaaa agacaggaaa ggtaattatt
     3481 ataggctctg gggtctcagg cttggcagca gctcgacagt tacaaagttt tggaatggat
     3541 gtcacacttt tggaagccag ggatcgtgtg ggtggacgag ttgccacatt tcgcaaagga
     3601 aactatgtag ctgatcttgg agccatggtg gtaacaggtc ttggagggaa tcctatggct
     3661 gtggtcagca aacaagtaaa tatggaactg gccaagatca agcaaaaatg cccactttat
     3721 gaagccaacg gacaagctgt tcctaaagag aaagatgaaa tggtagagca agagtttaac
     3781 cggttgctag aagctacatc ttaccttagt catcaactag acttcaatgt cctcaataat
     3841 aagcatgtgt cccttggcca ggcattggaa gttgtcattc agttacaaga gaagcatgtc
     3901 aaagatgagc agattgaaca ttggaagaag atagtgaaaa ctcaggaaga attgaaagaa
     3961 cttcttaata agatggtaaa tttgaaagag aaaattaaag aactccatca gcaatacaaa
     4021 gaagcatctg aagtaaagcc acccagagat attactgccg agttcttagt gaaaagcaaa
     4081 cacagggatc tgaccgccct atgcaaggaa tatgatgaat tagctgaaac acaaggaaag
     4141 ctagaagaaa aacttcagga gttggaagcg aatcccccaa gtgatgtata tctctcatca
     4201 agagacagac aaatacttga ttggcatttt gcaaatcttg aatttgctaa tgccacacct
     4261 ctctcaactc tctcccttaa gcactgggat caggatgatg actttgagtt cactggcagc
     4321 cacctgacag taaggaatgg ctactcgtgt gtgcctgtgg ctttagcaga aggcctagac
     4381 attaaactga atacagcagt gcgacaggtt cgctacacgg cttcaggatg tgaagtgata
     4441 gctgtgaata cccgctccac gagtcaaacc tttatttata aatgcgacgc agttctctgt
     4501 acccttcccc tgggtgtgct gaagcagcag ccaccagccg ttcagtttgt gccacctctc
     4561 cctgagtgga aaacatctgc agtccaaagg atgggatttg gcaaccttaa caaggtggtg
     4621 ttgtgttttg atcgggtgtt ctgggatcca agtgtcaatt tgttcgggca tgttggcagt
     4681 acgactgcca gcaggggtga gctcttcctc ttctggaacc tctataaagc tccaatactg
     4741 ttggcactag tggcaggaga agctgctggt atcatggaaa acataagtga cgatgtgatt
     4801 gttggccgat gcctggccat tctcaaaggg atttttggta gcagtgcagt acctcagccc
     4861 aaagaaactg tggtgtctcg ttggcgtgct gatccctggg ctcggggctc ttattcctat
     4921 gttgctgcag gatcatctgg aaatgactat gatttaatgg ctcagccaat cactcctggc
     4981 ccctcgattc caggtgcccc acagccgatt ccacgactct tctttgcggg agaacatacg
     5041 atccgtaact acccagccac agtgcatggt gctctgctga gtgggctgcg agaagcggga
     5101 agaattgcag accagttttt gggggccatg tatacgctgc ctcgccaggc cacaccaggt
     5161 gttcctgcac agcagtcccc aagcatgggc ggttccggcg gagggtcgat ggacaagaag
     5221 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
     5281 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
     5341 aagaacctga tcggcgccct gctgttcgac agcggagaaa cagccgaggc cacccggctg
     5401 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     5461 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     5521 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     5581 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     5641 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     5701 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     5761 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     5821 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     5881 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg
     5941 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     6001 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     6061 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     6121 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     6181 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     6241 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacatcgat
     6301 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     6361 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     6421 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     6481 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     6541 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     6601 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     6661 ggcgccagcg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     6721 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta caacgagctg
     6781 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     6841 aaaaaagcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     6901 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     6961 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     7021 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     7081 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     7141 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     7201 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     7261 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     7321 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     7381 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     7441 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     7501 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     7561 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     7621 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     7681 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag
     7741 agctttctga aggacgactc catcgataac aaagtgctga ctcggagcga caagaaccgg
     7801 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgc
     7861 cagctgctga atgccaagct gattacccag aggaagttcg acaatctgac caaggccgag
     7921 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     7981 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     8041 gagaacgaca aactgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     8101 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     8161 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     8221 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     8281 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     8341 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     8401 acaaacggcg aaacaggcga gatcgtgtgg gataagggcc gggactttgc caccgtgcgg
     8461 aaagtgctgt ctatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     8521 ttcagcaaag agtctatcct gcccaagagg aacagcgaca agctgatcgc cagaaagaag
     8581 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     8641 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     8701 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     8761 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     8821 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     8881 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     8941 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaaacac
     9001 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     9061 gctaatctgg acaaggtgct gagcgcctac aacaagcaca gagacaagcc tatcagagag
     9121 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     9181 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     9241 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     9301 ctgggaggcg acgcctatcc ctatgacgtg cccgattatg ccagcctggg cagcggctcc
     9361 cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc ggaaagtgga cggcattggt
     9421 agtgggagca acggcagcag cggatccgct actaacttca gcctgctgaa gcaggctgga
     9481 gacgtggagg agaaccctgg acctatggtg agcaagggcg aggaggataa catggccatc
     9541 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     9601 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     9661 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     9721 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     9781 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     9841 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     9901 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     9961 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
    10021 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
    10081 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
    10141 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
    10201 gagctgtaca agtaaggccg cgactctaga gtcgacctgc aggcatgcaa gcttgatatc
    10261 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
    10321 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
    10381 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
    10441 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
    10501 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
    10561 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
    10621 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
    10681 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
    10741 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
    10801 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
    10861 catcgatacc gtcgacctcg agggaattaa ttcgagctcg gtacctttaa gaccaatgac
    10921 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
    10981 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
    11041 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
    11101 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
    11161 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct agaattaatt
    11221 ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa
    11281 ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc tggcttactg
    11341 aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt tggacgaacc
    11401 ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc aatcagcagg
    11461 gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
    11521 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
    11581 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
    11641 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
    11701 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaatggtg
    11761 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
    11821 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
    11881 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
    11941 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
    12001 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
    12061 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
    12121 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
    12181 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
    12241 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
    12301 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
    12361 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
    12421 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
    12481 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
    12541 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
    12601 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
    12661 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
    12721 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
    12781 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
    12841 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
    12901 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
    12961 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
    13021 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
    13081 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    13141 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
    13201 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
    13261 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
    13321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
    13381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
    13441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
    13501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
    13561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
    13621 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
    13681 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
    13741 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
    13801 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
    13861 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
    13921 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
    13981 gattcattaa tgcagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
    14041 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
    14101 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
    14161 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
    14221 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
    14281 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
    14341 ggacacaaga caggcttgcg agatatgttt gagaatacca ctttatcccg cgtcagggag
    14401 aggcagtgcg taaaaagacg cggactcatg tgaaatactg gtttttagtg cgccagatct
    14461 ctataatctc gcgcaaccta ttttcccctc gaacactttt taagccgtag ataaacaggc
    14521 tgggacactt cacatgagcg aaaaatacat cgtcacctgg gacatgttgc agatccatgc
    14581 acgtaaactc gcaagccgac tgatgccttc tgaacaatgg aaaggcatta ttgccgtaag
    14641 ccgtggcggt ctgtaccggg tgcgttactg gcgcgtgaac tgggtattcg tcatgtcgat
    14701 accgtttgta tttccagcta cgatcacgac aaccagcgcg agcttaaagt gctgaaacgc
    14761 gcagaaggcg atggcgaagg cttcatcgtt attgatgacc tggtggatac cggtggtact
    14821 gcggttgcga ttcgtgaaat gtatccaaaa gcgcactttg tcaccatctt cgcaaaaccg
    14881 gctggtcgtc cgctggttga tgactatgtt gttgatatcc cgcaagatac ctggattgaa
    14941 cagccgtggg atatgggcgt cgtattcgtc ccgccaatct ccggtcgcta atcttttcaa
    15001 cgcctggcac tgccgggcgt tgttcttttt aacttcaggc gggttacaat agtttccagt
    15061 aagtattctg gaggctgcat ccatgacaca ggcaaacctg agcgaaaccc tgttcaaacc
    15121 ccgctttaaa catcctgaaa cctcgacgct agtccgccgc tttaatcacg gcgcacaacc
    15181 gcctgtgcag tcggcccttg atggtaaaac catccctcac tggtatcgca tgattaaccg
    15241 tctgatgtgg atctggcgcg gcattgaccc acgcgaaatc ctcgacgtcc aggcacgtat
    15301 tgtgatgagc gatgccgaac gtaccgacga tgatttatac gatacggtga ttggctaccg
    15361 tggcggcaac tggatttatg agtgggcccc ggatctttgt gaaggaacct tacttctgtg
    15421 gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat
    15481 ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa
    15541 cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt
    15601 gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc
    15661 ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt
    15721 ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa
    15781 aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa
    15841 gtaggcataa cagttataat cataacatac tgttttttct tactccacac aggcatagag
    15901 tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag
    15961 gggttaataa ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc
    16021 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
    16081 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
    16141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
    16201 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct
    16261 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg
    16321 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt
    16381 tgttaaatat gtactacaaa cttagtagt
//
