LOCUS       170_pB-Ef1a-CuO-       12373 bp DNA     circular SYN 16-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 12373)
  AUTHORS   Raquel Fueyo
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 2025 from SnapGene 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12373
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   137..171
                     /label=5'TR
                     /note="5'TR"
     primer_bind     complement(662..678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    686..702
                     /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(710..740)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    755..776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1064..1652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1823..2683)
                     /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"
     CDS             complement(1826..2683)
                     /label=Amp
                     /note="Amp"
     promoter        complement(2684..2788)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2814..3269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3411..3427
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3495..3514
                     /label=BII(R)
                     /note="BII(R)"
     repeat_region   3570..3600
                     /label=3'TR(1)
                     /note="3'TR(1)"
     promoter        4065..4276
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    4288..4315
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     regulatory      4333..4342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4369..4554
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             4555..4575
                     /codon_start=1
                     /standard_name="spacer"
                     /label=spacer
                     /note="krab(1)"
                     /note="krab(1);krab Linker"
                     /translation="GGSGGGS"
     CDS             4576..8679
                     /codon_start=1
                     /standard_name="dCas9 D10A&H840A (homo sapiens optimized
                     by MJ)"
                     /label=dCas9 D10A&H840A (homo sapiens optimized
                     by MJ)
                     /note="dCas9 D10A&H840A (homo sapiens optimized by MJ)"
                     /note="dCas9 D10A&H840A (homo sapiens optimized by 
                     MJ);dCas9;dCas9 D10A&H840A"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAGGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5183..8679
                     /codon_start=1
                     /standard_name="dCas9 D10A&H840A (homo sapiens optimized
                     by MJ)"
                     /label=dCas9 D10A&H840A (homo sapiens optimized
                     by MJ)
                     /note="dCas9 D10A&H840A (homo sapiens optimized by MJ)"
                     /note="dCas9 D10A&H840A (homo sapiens optimized by 
                     MJ);dCas9;dCas9 D10A&H840A"
                     /translation="GGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGL
                     TPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILR
                     VNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGG
                     ASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRR
                     QEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDK
                     GASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGE
                     QKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIK
                     DKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGR
                     LSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSL
                     HEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRE
                     RMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV
                     DAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKF
                     DNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVI
                     TLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKV
                     YDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVW
                     DKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGG
                     FDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKK
                     DLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED
                     NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII
                     HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
     mat_peptide     8650..8679
                     /standard_name="End of Cas9"
                     /label=End of Cas9
     CDS             join(8680..8709,8728..8748,8755..8814)
                     /codon_start=1
                     /standard_name="HA 2x NLS"
                     /label=HA 2x NLS
                     /note="HA 2x NLS"
                     /note="HA 2x NLS;HA 2x NLS;Feature edited by Mo Mandegar
                     This was done based on common features already included 
                     inSnapGene and the original annotated map by Stanley Qi 
                     andLuke Gilbert"
                     /translation="AYPYDVPDYAPKKKRKVPKKKRKVDGIGSGSNGSSGS"
     CDS             8683..8709
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8689..8715
                     /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             8728..8748
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8755..8775
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8815..8871
                     /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             8872..9573
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             8872..9572
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=monomeric blue fluorescent protein
                     /note="TagBFP"
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(9548..9581)
                     /label=Primer 2
     promoter        9598..9809
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             9822..10090
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      10118..10127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10130..10156
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10136..10162
                     /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             10274..10882
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     CDS             10919..10972
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    10979..11579
                     /label=Puro
     misc_feature    11626..11756
                     /label=SV40 pA
                     /note="SV40 pA"
     polyA_signal    11626..11747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        complement(12005..12235)
                     /note="core insul(1)"
ORIGIN
        1 ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa
       61 gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag
      121 caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaatctag
      181 ctgcatcagg atcatatcgt cgggtctttt ttccggctca gtcatcgccc aagctggcgc
      241 tatctgggca tcggggagga agaagcccgt gccttttccc gcgaggttga agcggcatgg
      301 aaagagtttg ccgaggatga ctgctgctgc attgacgttg agcgaaaacg cacgtttacc
      361 atgatgattc gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt cgttcaggcc
      421 acctgggata ccagttcgtc gcggcttttc cggacacagt tccggatggt cagcccgaag
      481 cgcatcagca acccgaacaa taccggcgac agccggaact gccgtgccgg tgtgcagatt
      541 aatgacagcg gtgcggcgct gggatattac gtcagcgagg acgggtatcc tggctggatg
      601 ccgcagaaat ggacatggat accccgtgag ttacccggcg ggcgcgcttg gcgtaatcat
      661 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      721 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      781 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      841 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
      901 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1501 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1621 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1681 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1741 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1801 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1861 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1921 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1981 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2041 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2101 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2161 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2221 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2281 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2341 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2401 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2461 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2521 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2581 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2641 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2701 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2761 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     2821 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2881 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2941 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3001 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3061 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3121 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3181 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3241 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     3301 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     3361 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     3421 ggccagtgag cgcgcctcgt tcattcacgt ttttgaaccc gtggaggacg ggcagactcg
     3481 cggtgcaaat gtgttttaca gcgtgatgga gcagatgaag atgctcgaca cgctgcagaa
     3541 cacgcagcta gattaaccct agaaagataa tcatattgtg acgtacgtta aagataatca
     3601 tgtgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg
     3661 aatagatatt aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa
     3721 tttatttatg tttatttatt tattaaaaaa aacaaaaact caaaatttct tctataaagt
     3781 aacaaaactt ttatgaggga cagccccccc cccaaagccc ccagggatgt aattacgtcc
     3841 ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc
     3901 ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga
     3961 tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg
     4021 gggaaaaggc ctccacggcc agactagttc cggtgcccgt cagtgggcag agcgcacatc
     4081 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag
     4141 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     4201 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     4261 tgccgccaga acacagcccc cggtatatac aaacagacca gattgtctgt ttgtttttgg
     4321 tctagagcta gcgccaccat ggatgctaag tcactaactg cctggtcccg gacactggtg
     4381 accttcaagg atgtatttgt ggacttcacc agggaggagt ggaagctgct ggacactgct
     4441 cagcagatcg tgtacagaaa tgtgatgctg gagaactata agaacctggt ttccttgggt
     4501 tatcagctta ctaagccaga tgtgatcctc cggttggaga agggagaaga gcccggcggt
     4561 tccggcggag ggtcgatgga caagaagtac agcatcggcc tggccatcgg caccaactct
     4621 gtaggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
     4681 ggcaacaccg accggcacag catcaagaag aacctgatcg gcgccctgct gttcgacagc
     4741 ggagaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
     4801 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
     4861 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     4921 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     4981 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     5041 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     5101 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     5161 ttcgaggaaa accccatcaa cgccggcggc gtggacgcca aggccatcct gtctgccaga
     5221 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     5281 ggcctgttcg gcaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     5341 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     5401 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     5461 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     5521 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     5581 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     5641 aagaacggct acgccggcta catcgatggc ggagccagcc aggaagagtt ctacaagttc
     5701 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     5761 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     5821 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     5881 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     5941 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     6001 tggaacttcg aggaagtggt ggacaagggc gccagcgccc agagcttcat cgagcggatg
     6061 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     6121 gagtacttca ccgtgtacaa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     6181 aagcccgcct tcctgagcgg cgagcagaaa aaagccatcg tggacctgct gttcaagacc
     6241 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     6301 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     6361 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     6421 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     6481 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     6541 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     6601 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     6661 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     6721 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     6781 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     6841 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     6901 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     6961 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     7021 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     7081 tacgatgtgg acgctatcgt gcctcagagc tttctgaagg acgactccat cgataacaaa
     7141 gtgctgactc ggagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     7201 gtgaagaaga tgaagaacta ctggcgccag ctgctgaatg ccaagctgat tacccagagg
     7261 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     7321 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     7381 gactcccgga tgaacactaa gtacgacgag aacgacaaac tgatccggga agtgaaagtg
     7441 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     7501 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     7561 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     7621 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     7681 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     7741 gagatccgga agcggcctct gatcgagaca aacggcgaaa caggcgagat cgtgtgggat
     7801 aagggccggg actttgccac cgtgcggaaa gtgctgtcta tgccccaagt gaatatcgtg
     7861 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     7921 agcgacaagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     7981 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     8041 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     8101 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     8161 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     8221 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     8281 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     8341 cagctgtttg tggaacagca caaacactac ctggacgaga tcatcgagca gatcagcgag
     8401 ttctccaaga gagtgatcct ggccgacgct aatctggaca aggtgctgag cgcctacaac
     8461 aagcacagag acaagcctat cagagagcag gccgagaata tcatccacct gtttaccctg
     8521 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     8581 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     8641 tacgagacac ggatcgacct gtctcagctg ggaggcgacg cctatcccta tgacgtgccc
     8701 gattatgcca gcctgggcag cggctccccc aagaaaaaac gcaaggtgga agatcctaag
     8761 aaaaagcgga aagtggacgg cattggtagt gggagcaacg gcagcagcgg atccgctact
     8821 aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc tatgagcgag
     8881 ctgattaagg agaacatgca catgaagctg tacatggagg gcaccgtgga caaccatcac
     8941 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc
     9001 aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac tagcttcctc
     9061 tacggcagca agaccttcat caaccacacc cagggcatcc ccgacttctt caagcagtcc
     9121 ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc
     9181 gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg
     9241 gtgaacttca catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccttc
     9301 accgagacgc tgtaccccgc tgacggcggc ctggaaggca gaaacgacat ggccctgaag
     9361 ctcgtgggcg ggagccatct gatcgcaaac atcaagacca catatagatc caagaaaccc
     9421 gctaagaacc tcaagatgcc tggcgtctac tatgtggact acagactgga aagaatcaag
     9481 gaggccaaca acgagaccta cgtcgagcag cacgaggtgg cagtggccag atactgcgac
     9541 ctccctagca aactggggca caagcttaat taagcgatcg ctccggtgcc cgtcagtggg
     9601 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg
     9661 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     9721 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     9781 ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt
     9841 cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc
     9901 ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt
     9961 cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca
    10021 cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc
    10081 gttacagatc caagctgtga ccggcgccta ctctagagcc gccatggctt acccatacga
    10141 tgttccagat tacgctagtt tggtgaggca gcagagaccg atgacatcca ctttgccttt
    10201 ctctccacag gtgtccactc ccaggtccaa gtttggaaga tctaccatgc caaagagacc
    10261 cagacccgtg atcatgagtc caaagagaag aacacaggca gagcgcgcaa tggagaccca
    10321 gggcaagttg attgcagcgg ccctgggggt tttacgggaa aaaggttacg cgggattccg
    10381 gatcgcagat gtgcccggtg ctgcaggtgt ctcgagagga gcgcagagcc atcatttccc
    10441 gacaaagctt gagcttctgc ttgccacttt tgaatggctt tacgaacaga tcaccgaacg
    10501 cagtcgggct cgattagcga aattgaagcc agaggatgac gtcatccagc aaatgctgga
    10561 cgacgccgcc gaatttttcc tcgacgatga cttctctatc agccttgatt tgattgtggc
    10621 tgccgaccgg gatccagcgt tacgcgaggg tattcagcgc acggtagaga ggaatcggtt
    10681 tgtcgtcgag gatatgtggc ttggtgttct ggtgagccgt ggtctttcgc gtgatgatgc
    10741 agaagatatc ctttggttga tattcaattc ggtgcgtggg cttgctgttc gtagcctatg
    10801 gcagaaggac aaagaacgct ttgagcgtgt caggaactcg acactcgaaa ttgcgcgaga
    10861 gcggtacgcg aaattcaagc gcgcggccaa accctctggc cggtctctct ctgttcgaga
    10921 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctgaattcat
    10981 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
    11041 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
    11101 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
    11161 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
    11221 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
    11281 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
    11341 cgcgtggttc ccggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
    11401 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctcgg
    11461 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
    11521 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
    11581 tcgacaatca acctctggat tacaaaattt gtggttaact aactaaactt gtttattgca
    11641 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    11701 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggaat
    11761 tgactcaaat gatgtcaatt agtctatcag aagctcatct ggtctccctt ccgggggaca
    11821 agacatccct gtttaatatt taaacagcag tgttcccaaa ctgggttctt atatcccttg
    11881 ctctggtcaa ccaggttgca gggtttcctg tcctcacagg aacgaagtcc ctaaagaaac
    11941 agtggcagcc aggtttagcc ccggaattga ctggattcct tttttagggc ccattggtat
    12001 ggctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca
    12061 gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct
    12121 cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc
    12181 tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg tccctgatat
    12241 ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat
    12301 ataattatcg tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg
    12361 actcacgcgg tcg
//
