LOCUS       sequence-287200-       16411 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 16411)
  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 16411)
  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..16411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..17)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2..18)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             108..341
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(366..384)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(367..384)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(674..693)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(907..926)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1026..1048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1086..1104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1172..1276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1277..1345,1346..2137)
                     /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(1495..1514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2308..2896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2797..2816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3050..3067
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3137..3157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3142..3471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3322..3457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4605..4670
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4800..4820
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5245..5379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5282..5301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5336..5355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5548..6181
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6228..6353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6850..7082
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7267..7311
                     /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    7596..7713
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7775..8124
                     /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    7783..7801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7819..7837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7855..7873
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7891..7909
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7927..7945
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7963..7981
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7999..8017
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     8063..8087
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(10755..10781,10782..10784,10785..13271,13272..13274,
                     13275..14858)
                     /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             10755..14858
                     /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             14862..14888
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             14868..14894
                     /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             14907..14927
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14934..14954
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14994..15050
                     /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             15051..15761
                     /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(15196..15214)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     15464..15483
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    15821..16409
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(15874..15894)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(16292..16303)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 cgataccgtc gacctcgagg gaattaattc gagctcggta cctttaagac caatgactta
       61 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
      121 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
      181 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
      241 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
      301 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatctaga attaattccg
      361 tgtattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg
      421 tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg tagcatctgg cttactgaag
      481 cagaccctat catctctctc gtaaactgcc gtcagagtcg gtttggttgg acgaaccttc
      541 tgagtttctg gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat cagcagggtc
      601 atcgctagcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
      661 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
      721 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
      781 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
      841 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg aatggtgcac
      901 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
      961 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1021 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     1081 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1141 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1201 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1261 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1321 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1381 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1441 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1501 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1561 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1621 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1681 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1741 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1801 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1861 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1921 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1981 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2041 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2101 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2161 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2221 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2281 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2341 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2401 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtctttct
     2461 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2521 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2581 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2641 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2701 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2761 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2821 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2881 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2941 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3001 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3061 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3121 tcattaatgc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgga
     3481 cacaagacag gcttgcgaga tatgtttgag aataccactt tatcccgcgt cagggagagg
     3541 cagtgcgtaa aaagacgcgg actcatgtga aatactggtt tttagtgcgc cagatctcta
     3601 taatctcgcg caacctattt tcccctcgaa cactttttaa gccgtagata aacaggctgg
     3661 gacacttcac atgagcgaaa aatacatcgt cacctgggac atgttgcaga tccatgcacg
     3721 taaactcgca agccgactga tgccttctga acaatggaaa ggcattattg ccgtaagccg
     3781 tggcggtctg taccgggtgc gttactggcg cgtgaactgg gtattcgtca tgtcgatacc
     3841 gtttgtattt ccagctacga tcacgacaac cagcgcgagc ttaaagtgct gaaacgcgca
     3901 gaaggcgatg gcgaaggctt catcgttatt gatgacctgg tggataccgg tggtactgcg
     3961 gttgcgattc gtgaaatgta tccaaaagcg cactttgtca ccatcttcgc aaaaccggct
     4021 ggtcgtccgc tggttgatga ctatgttgtt gatatcccgc aagatacctg gattgaacag
     4081 ccgtgggata tgggcgtcgt attcgtcccg ccaatctccg gtcgctaatc ttttcaacgc
     4141 ctggcactgc cgggcgttgt tctttttaac ttcaggcggg ttacaatagt ttccagtaag
     4201 tattctggag gctgcatcca tgacacaggc aaacctgagc gaaaccctgt tcaaaccccg
     4261 ctttaaacat cctgaaacct cgacgctagt ccgccgcttt aatcacggcg cacaaccgcc
     4321 tgtgcagtcg gcccttgatg gtaaaaccat ccctcactgg tatcgcatga ttaaccgtct
     4381 gatgtggatc tggcgcggca ttgacccacg cgaaatcctc gacgtccagg cacgtattgt
     4441 gatgagcgat gccgaacgta ccgacgatga tttatacgat acggtgattg gctaccgtgg
     4501 cggcaactgg atttatgagt gggccccgga tctttgtgaa ggaaccttac ttctgtggtg
     4561 tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt
     4621 taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct
     4681 atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct
     4741 cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc
     4801 caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt
     4861 tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac accacaaagg
     4921 aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc tttataagta
     4981 ggcataacag ttataatcat aacatactgt tttttcttac tccacacagg catagagtgt
     5041 ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt tgtaaagggg
     5101 ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag ccataccaca
     5161 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5221 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5341 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     5401 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctagtggtt
     5461 tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg
     5521 ttaaatatgt actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag
     5581 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     5641 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     5701 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     5761 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     5821 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     5881 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     5941 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     6001 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6061 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     6241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     6301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     6361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     6421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     6481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     6541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     6601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     6661 aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg
     6721 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     6781 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     6841 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     6901 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     6961 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     7021 catcaagcag ctccaggcaa gaatcctggc tgtgaaagat acctaaagga tcaacagctc
     7081 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     7141 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     7201 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     7261 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     7321 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     7381 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     7441 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     7501 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     7561 cgacggtatc gccaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     7621 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     7681 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     7741 ccagtttatc gatgaggccc tttcgtcttc actcgagttt actccctatc agtgatagag
     7801 aacgtatgaa gagtttactc cctatcagtg atagagaacg tatgcagact ttactcccta
     7861 tcagtgatag agaacgtata aggagtttac tccctatcag tgatagagaa cgtatgacca
     7921 gtttactccc tatcagtgat agagaacgta tctacagttt actccctatc agtgatagag
     7981 aacgtatatc cagtttactc cctatcagtg atagagaacg tatgtcgagg taggcgtgta
     8041 cggtgggcgc ctataaaagc agagctcgtt tagtgaaccg tcagatcgcc tggagcaatt
     8101 ccacaacact tttgtcttat acttacgcgt ctgcaggata tcaagcttgc ggtaccgcgg
     8161 gcccgggatc cgccaccatg ttatctggga agaaggcggc agccgcggcg gcggcggctg
     8221 cagcggcagc aaccgggacg gaggctggcc ctgggacagc aggcggctcc gagaacgggt
     8281 ctgaggtggc cgcgcagccc gcgggcctgt cgggcccagc cgaggtcggg ccgggggcgg
     8341 tgggggagcg cacaccccgc aagaaagagc ctccgcgggc ctcgcccccc gggggcctgg
     8401 cggaaccgcc ggggtccgca gggcctcagg ccggccctac tgtcgtgcct gggtctgcga
     8461 cccccatgga aactggaata gcagagactc cggaggggcg tcggaccagc cggcgcaagc
     8521 gggcgaaggt agagtacaga gagatggatg aaagcttggc caacctctca gaagatgagt
     8581 attattcaga agaagagaga aatgccaaag cagagaagga aaagaagctt cccccaccac
     8641 cccctcaagc cccacctgag gaagaaaatg aaagtgagcc tgaagaacca tcgggtgtgg
     8701 agggcgcagc tttccagagc cgacttcctc atgaccggat gacttctcaa gaagcagcct
     8761 gttttccaga tattatcagt ggaccacaac agacccagaa ggtttttctt ttcattagaa
     8821 accgcacact gcagttgtgg ttggataatc caaagattca gctgacattt gaggctactc
     8881 tccaacaatt agaagcacct tataacagtg atactgtgct tgtccaccga gttcacagtt
     8941 atttagagcg tcatggtctt atcaacttcg gcatctataa gaggataaaa cccctaccaa
     9001 ctaaaaagac aggaaaggta attattatag gctctggggt ctcaggcttg gcagcagctc
     9061 gacagttaca aagttttgga atggatgtca cacttttgga agccagggat cgtgtgggtg
     9121 gacgagttgc cacatttcgc aaaggaaact atgtagctga tcttggagcc atggtggtaa
     9181 caggtcttgg agggaatcct atggctgtgg tcagcaaaca agtaaatatg gaactggcca
     9241 agatcaagca aaaatgccca ctttatgaag ccaacggaca agctgttcct aaagagaaag
     9301 atgaaatggt agagcaagag tttaaccggt tgctagaagc tacatcttac cttagtcatc
     9361 aactagactt caatgtcctc aataataagc atgtgtccct tggccaggca ttggaagttg
     9421 tcattcagtt acaagagaag catgtcaaag atgagcagat tgaacattgg aagaagatag
     9481 tgaaaactca ggaagaattg aaagaacttc ttaataagat ggtaaatttg aaagagaaaa
     9541 ttaaagaact ccatcagcaa tacaaagaag catctgaagt aaagccaccc agagatatta
     9601 ctgccgagtt cttagtgaaa agcaaacaca gggatctgac cgccctatgc aaggaatatg
     9661 atgaattagc tgaaacacaa ggaaagctag aagaaaaact tcaggagttg gaagcgaatc
     9721 ccccaagtga tgtatatctc tcatcaagag acagacaaat acttgattgg cattttgcaa
     9781 atcttgaatt tgctaatgcc acacctctct caactctctc ccttaagcac tgggatcagg
     9841 atgatgactt tgagttcact ggcagccacc tgacagtaag gaatggctac tcgtgtgtgc
     9901 ctgtggcttt agcagaaggc ctagacatta aactgaatac agcagtgcga caggttcgct
     9961 acacggcttc aggatgtgaa gtgatagctg tgaatacccg ctccacgagt caaaccttta
    10021 tttataaatg cgacgcagtt ctctgtaccc ttcccctggg tgtgctgaag cagcagccac
    10081 cagccgttca gtttgtgcca cctctccctg agtggaaaac atctgcagtc caaaggatgg
    10141 gatttggcaa ccttaacaag gtggtgttgt gttttgatcg ggtgttctgg gatccaagtg
    10201 tcaatttgtt cgggcatgtt ggcagtacga ctgccagcag gggtgagctc ttcctcttct
    10261 ggaacctcta taaagctcca atactgttgg cactagtggc aggagaagct gctggtatca
    10321 tggaaaacat aagtgacgat gtgattgttg gccgatgcct ggccattctc aaagggattt
    10381 ttggtagcag tgcagtacct cagcccaaag aaactgtggt gtctcgttgg cgtgctgatc
    10441 cctgggctcg gggctcttat tcctatgttg ctgcaggatc atctggaaat gactatgatt
    10501 taatggctca gccaatcact cctggcccct cgattccagg tgccccacag ccgattccac
    10561 gactcttctt tgcgggagaa catacgatcc gtaactaccc agccacagtg catggtgctc
    10621 tgctgagtgg gctgcgagaa gcgggaagaa ttgcagacca gtttttgggg gccatgtata
    10681 cgctgcctcg ccaggccaca ccaggtgttc ctgcacagca gtccccaagc atgggcggtt
    10741 ccggcggagg gtcgatggac aagaagtaca gcatcggcct ggccatcggc accaactctg
    10801 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
    10861 gcaacaccga ccggcacagc atcaagaaga acctgatcgg cgccctgctg ttcgacagcg
    10921 gagaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
    10981 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
    11041 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
    11101 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
    11161 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
    11221 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
    11281 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
    11341 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
    11401 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
    11461 gcctgttcgg caacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
    11521 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
    11581 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
    11641 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
    11701 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
    11761 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
    11821 agaacggcta cgccggctac atcgatggcg gagccagcca ggaagagttc tacaagttca
    11881 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
    11941 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
    12001 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
    12061 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
    12121 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
    12181 ggaacttcga ggaagtggtg gacaagggcg ccagcgccca gagcttcatc gagcggatga
    12241 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
    12301 agtacttcac cgtgtacaac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
    12361 agcccgcctt cctgagcggc gagcagaaaa aagccatcgt ggacctgctg ttcaagacca
    12421 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
    12481 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
    12541 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
    12601 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
    12661 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
    12721 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
    12781 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
    12841 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
    12901 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
    12961 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
    13021 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
    13081 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
    13141 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
    13201 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
    13261 acgatgtgga cgctatcgtg cctcagagct ttctgaagga cgactccatc gataacaaag
    13321 tgctgactcg gagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
    13381 tgaagaagat gaagaactac tggcgccagc tgctgaatgc caagctgatt acccagagga
    13441 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
    13501 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
    13561 actcccggat gaacactaag tacgacgaga acgacaaact gatccgggaa gtgaaagtga
    13621 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
    13681 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
    13741 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
    13801 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
    13861 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
    13921 agatccggaa gcggcctctg atcgagacaa acggcgaaac aggcgagatc gtgtgggata
    13981 agggccggga ctttgccacc gtgcggaaag tgctgtctat gccccaagtg aatatcgtga
    14041 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
    14101 gcgacaagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
    14161 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
    14221 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
    14281 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
    14341 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
    14401 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
    14461 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
    14521 agctgtttgt ggaacagcac aaacactacc tggacgagat catcgagcag atcagcgagt
    14581 tctccaagag agtgatcctg gccgacgcta atctggacaa ggtgctgagc gcctacaaca
    14641 agcacagaga caagcctatc agagagcagg ccgagaatat catccacctg tttaccctga
    14701 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
    14761 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
    14821 acgagacacg gatcgacctg tctcagctgg gaggcgacgc ctatccctat gacgtgcccg
    14881 attatgccag cctgggcagc ggctccccca agaaaaaacg caaggtggaa gatcctaaga
    14941 aaaagcggaa agtggacggc attggtagtg ggagcaacgg cagcagcgga tccgctacta
    15001 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct atggtgagca
    15061 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
    15121 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
    15181 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
    15241 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
    15301 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
    15361 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
    15421 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
    15481 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
    15541 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
    15601 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
    15661 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
    15721 gccgccactc caccggcggc atggacgagc tgtacaagta agcggccgcg actctagagt
    15781 cgacctgcag gcatgcaagc ttgatatcaa gcttatcgat aatcaacctc tggattacaa
    15841 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
    15901 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
    15961 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
    16021 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
    16081 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
    16141 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
    16201 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
    16261 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
    16321 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
    16381 tcggatctcc ctttgggccg cctccccgca t
//
