LOCUS       sequence-181086-       14133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid expressing dCas9-2xAM and gRNA for human IRF-1 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14133)
  AUTHORS   Fujita T, Yuno M, Fujii H
  TITLE     enChIP systems using different CRISPR orthologues and epitope tags.
  JOURNAL   BMC Res Notes. 2018 Feb 27;11(1):154. doi: 
            10.1186/s13104-018-3262-4.
  PUBMED    29482606
REFERENCE   2  (bases 1 to 14133)
  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: "BMC Res 
            Notes."; date: "2018-02-27"; volume: "11(1):154. doi"; pages: " 
            10.1186/s13104-018-3262-4"
COMMENT     Sequence Label: pLenti_dCas9-2xAM_hIRF-1
FEATURES             Location/Qualifiers
     source          1..14133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          127..354
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      366..375
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(372..398,399..401,402..2885,2886..2891,2892..2957,
                     2958..2960,2961..4475)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /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             join(372..398,399..401,402..2888,2889..2891,2892..4475)
                     /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             372..4475
                     /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             4488..4508
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4614..4670
                     /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             4671..5270
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4671..4690)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5167..5187
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5300..5888
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5353..5373)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5771..5782)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6407..6587
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6613..6630)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6619..6843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6889..7317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6976..6995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7186..7207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7326..7346)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7331..7660
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7511..7646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7573..7592
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7708..7755
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7774..8148
                     /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    8278..8399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8315..8334)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8369..8388
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8448..8464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8448..8464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8461..8483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8472..8488
                     /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(8496..8502,8503..8520,8521..8526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8541..8562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8679..8696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8850..9438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8930..8949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9609..10400,10401..10469))
                     /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     10232..10251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10470..10574)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10649..10668)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10840..11219
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11221..11419
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11374..11394
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11437..11617
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11664..11789
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12281..12514
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12699..12743
                     /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    13017..13134
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        13273..13521
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     13273..13293
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     13444..13463
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        13542..13617
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        13714..13999
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
       61 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      121 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      181 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      241 tctcctccgg gctgtaatta gctgagcaag aggtaagggt ttaagggatg gttggttggt
      301 ggggtattaa tgtttaatta cctggagcac ctgcctgaaa tcactttttt tcaggttgga
      361 ccggtgccac catggacaag aagtactcca ttgggctcgc tatcggcaca aacagcgtcg
      421 gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa gttctgggca
      481 ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc gactccgggg
      541 agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga
      601 atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg gatgactctt
      661 tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac gagcgccacc
      721 caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca accatatatc
      781 atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg atctatctcg
      841 cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac ctgaacccag
      901 acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat cagcttttcg
      961 aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc gctaggctgt
     1021 ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag aagaacggcc
     1081 tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa tctaacttcg
     1141 acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat gatctcgaca
     1201 atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca aagaacctgt
     1261 cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc aaagctccgc
     1321 tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact ttgctgaagg
     1381 cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat cagtctaaaa
     1441 atggctacgc cggatacatt gacggcggag caagccagga ggaattttac aaatttatta
     1501 agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt aacagagaag
     1561 atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag attcacctgg
     1621 gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg aaagataaca
     1681 gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc cccctcgccc
     1741 ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc actccctgga
     1801 acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa aggatgacta
     1861 actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg ctgtacgagt
     1921 acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc
     1981 cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc aagacgaacc
     2041 ggaaagttac cgtgaaacag ctcaaagagg actatttcaa aaagattgaa tgtttcgact
     2101 ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg tatcacgatc
     2161 tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag gacattcttg
     2221 aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa gaacgcttga
     2281 aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg cgccgatata
     2341 caggatgggg gcggctgtca agaaaactga tcaatgggat cagagacaag cagagtggaa
     2401 agacaatcct ggattttctt aagtccgatg gatttgccaa caggaacttc atgcagttga
     2461 tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt tctggccagg
     2521 gggacagtct gcacgagcac atcgctaatc ttgcaggtag cccagctatc aaaaagggaa
     2581 tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg cataagcccg
     2641 agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga cagaagaaca
     2701 gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc caaatcctta
     2761 aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg tactacctgc
     2821 agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc tccgactacg
     2881 acgtggatgc catcgtgccc cagtcttttc tcaaagatga ttctattgat aataaagtgt
     2941 tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa gaagttgtca
     3001 agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca caacggaagt
     3061 tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa gccggcttca
     3121 tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa attctcgatt
     3181 cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg aaagttatta
     3241 ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat aaggtgagag
     3301 agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta ggcactgcac
     3361 ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat aaagtgtacg
     3421 atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc gctaagtact
     3481 tcttttacag caatattatg aattttttca agaccgagat tacactggcc aatggagaga
     3541 ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg
     3601 gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa
     3661 agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa aggaacagcg
     3721 acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga ttcgattctc
     3781 ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac
     3841 tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa
     3901 accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac ctcatcatta
     3961 agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg
     4021 cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt aatttcttgt
     4081 atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag cagaagcagc
     4141 tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata agcgaattct
     4201 ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct tacaataagc
     4261 acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt actctgacca
     4321 acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga aagcggtaca
     4381 cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg gggctctatg
     4441 aaacaagaat cgacctctct cagctcggtg gagacagcag ggctgacccc aagaagaaga
     4501 ggaaggtggg gcaagatcct caacgcaaag gcaacgtgat actctctcag gcttacgggc
     4561 aagatcctca acgcaaaggc aacgtgatac tctctcaggc ttacggatcc ggagctacta
     4621 acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct atgaccgagt
     4681 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4741 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4801 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4861 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4921 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4981 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     5041 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     5101 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     5161 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     5221 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gttaacatcg
     5281 agggatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5341 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5401 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5461 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5521 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5581 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5641 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     5701 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     5761 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5821 gcggcctctt ccgcgtattc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     5881 ctccccgcat cgataccgtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     5941 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     6001 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6061 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gatgacaaga
     6121 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
     6181 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     6241 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
     6301 gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg
     6361 cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt
     6421 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6481 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6541 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa
     6601 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6661 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6721 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6781 acagcaaggg ggaggattgg gatgacaata gcaggcatgc tggggatgcg gtgggctcta
     6841 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     6901 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6961 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     7021 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     7081 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7141 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7201 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     7261 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     7321 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     7381 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     7441 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     7501 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     7561 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     7621 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     7681 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     7741 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     7801 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     7861 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     7921 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     7981 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     8041 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     8101 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     8161 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     8221 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     8281 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     8341 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     8401 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     8461 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8521 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8581 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8641 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8701 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8761 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     8821 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     8881 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8941 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     9001 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     9061 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     9121 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     9181 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     9241 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9301 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9361 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9421 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9481 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9541 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9601 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9661 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9721 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9781 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     9841 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9901 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9961 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
    10021 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
    10081 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    10141 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    10201 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    10261 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10321 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10381 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10441 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10501 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10561 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
    10621 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
    10681 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
    10741 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
    10801 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
    10861 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
    10921 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
    10981 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
    11041 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
    11101 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
    11161 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
    11221 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
    11281 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
    11341 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
    11401 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
    11461 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
    11521 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
    11581 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
    11641 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
    11701 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
    11761 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
    11821 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
    11881 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
    11941 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
    12001 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
    12061 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
    12121 agcaagggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
    12181 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
    12241 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
    12301 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
    12361 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
    12421 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
    12481 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
    12541 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
    12601 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
    12661 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
    12721 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
    12781 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
    12841 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
    12901 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
    12961 gagacagatc cattcgatta gtgaacggat ctacaaatgg cagtattcat ccacaatttt
    13021 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
    13081 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
    13141 tattacaggg acagcagaaa ttcactttga attaattcaa gcttcgtgag gctccggtgc
    13201 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
    13261 caattgaacc gggagggcct atttcccatg attccttcat atttgcatat acgatacaag
    13321 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
    13381 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
    13441 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
    13501 ttgtggaaag gacgaaacac cgcgggggcg ctgggctgtc cgttttagag ctagaaatag
    13561 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
    13621 tttgttttag agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg
    13681 ccaattctgc agacaaatgg ctctagaggt acccgttaca taacttacgg taaatggccc
    13741 gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc caatagggac
    13801 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
    13861 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
    13921 gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
    13981 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
    14041 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
    14101 gggggcgggg ggggggggcg cgcgcgcnag gcg
//
