LOCUS       sequence-68222-e        7785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mammalian cell codon-optimized dCas9-VP64.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7785)
  AUTHORS   Maeder ML, Linder SJ, Cascio VM, Fu Y, Ho QH, Joung JK
  TITLE     CRISPR RNA-guided activation of endogenous human genes.
  JOURNAL   Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2598.
  PUBMED    23892898
REFERENCE   2  (bases 1 to 7785)
  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 
            Methods. 2013 Jul 25. doi: 10.1038/nmeth.2598."
COMMENT     Sequence Label: pMLM3705
FEATURES             Location/Qualifiers
     source          1..7785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(409..3810,3811..3813,3814..4410,4411,4412..4414,
                     4415..4416,4417..4419,4420..4512)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /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(409..435,436..438,439..2925,2926..2928,2929..4512)
                     /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             4519..4539
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4546..4611
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4642..4791
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             4800..4817
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4840..4857)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4846..5070
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5141..5157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5141..5157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5154..5176)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5165..5181
                     /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(5189..5195,5196..5213,5214..5219))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5234..5255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5372..5389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5543..6131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5623..5642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6302..7093,7094..7162))
                     /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     6925..6944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7163..7267)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7346..7365)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7537..7785,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggataaaaag
      421 tattctattg gtttagccat cggcactaat tccgttggat gggctgtcat aaccgatgaa
      481 tacaaagtac cttcaaagaa atttaaggtg ttggggaaca cagaccgtca ttcgattaaa
      541 aagaatctta tcggtgccct cctattcgat agtggcgaaa cggcagaggc gactcgcctg
      601 aaacgaaccg ctcggagaag gtatacacgt cgcaagaacc gaatatgtta cttacaagaa
      661 atttttagca atgagatggc caaagttgac gattctttct ttcaccgttt ggaagagtcc
      721 ttccttgtcg aagaggacaa gaaacatgaa cggcacccca tctttggaaa catagtagat
      781 gaggtggcat atcatgaaaa gtacccaacg atttatcacc tcagaaaaaa gctagttgac
      841 tcaactgata aagcggacct gaggttaatc tacttggctc ttgcccatat gataaagttc
      901 cgtgggcact ttctcattga gggtgatcta aatccggaca actcggatgt cgacaaactg
      961 ttcatccagt tagtacaaac ctataatcag ttgtttgaag agaaccctat aaatgcaagt
     1021 ggcgtggatg cgaaggctat tcttagcgcc cgcctctcta aatcccgacg gctagaaaac
     1081 ctgatcgcac aattacccgg agagaagaaa aatgggttgt tcggtaacct tatagcgctc
     1141 tcactaggcc tgacaccaaa ttttaagtcg aacttcgact tagctgaaga tgccaaattg
     1201 cagcttagta aggacacgta cgatgacgat ctcgacaatc tactggcaca aattggagat
     1261 cagtatgcgg acttattttt ggctgccaaa aaccttagcg atgcaatcct cctatctgac
     1321 atactgagag ttaatactga gattaccaag gcgccgttat ccgcttcaat gatcaaaagg
     1381 tacgatgaac atcaccaaga cttgacactt ctcaaggccc tagtccgtca gcaactgcct
     1441 gagaaatata aggaaatatt ctttgatcag tcgaaaaacg ggtacgcagg ttatattgac
     1501 ggcggagcga gtcaagagga attctacaag tttatcaaac ccatattaga gaagatggat
     1561 gggacggaag agttgcttgt aaaactcaat cgcgaagatc tactgcgaaa gcagcggact
     1621 ttcgacaacg gtagcattcc acatcaaatc cacttaggcg aattgcatgc tatacttaga
     1681 aggcaggagg atttttatcc gttcctcaaa gacaatcgtg aaaagattga gaaaatccta
     1741 acctttcgca taccttacta tgtgggaccc ctggcccgag ggaactctcg gttcgcatgg
     1801 atgacaagaa agtccgaaga aacgattact ccatggaatt ttgaggaagt tgtcgataaa
     1861 ggtgcgtcag ctcaatcgtt catcgagagg atgaccaact ttgacaagaa tttaccgaac
     1921 gaaaaagtat tgcctaagca cagtttactt tacgagtatt tcacagtgta caatgaactc
     1981 acgaaagtta agtatgtcac tgagggcatg cgtaaacccg cctttctaag cggagaacag
     2041 aagaaagcaa tagtagatct gttattcaag accaaccgca aagtgacagt taagcaattg
     2101 aaagaggact actttaagaa aattgaatgc ttcgattctg tcgagatctc cggggtagaa
     2161 gatcgattta atgcgtcact tggtacgtat catgacctcc taaagataat taaagataag
     2221 gacttcctgg ataacgaaga gaatgaagat atcttagaag atatagtgtt gactcttacc
     2281 ctctttgaag atcgggaaat gattgaggaa agactaaaaa catacgctca cctgttcgac
     2341 gataaggtta tgaaacagtt aaagaggcgt cgctatacgg gctggggacg attgtcgcgg
     2401 aaacttatca acgggataag agacaagcaa agtggtaaaa ctattctcga ttttctaaag
     2461 agcgacggct tcgccaatag gaactttatg cagctgatcc atgatgactc tttaaccttc
     2521 aaagaggata tacaaaaggc acaggtttcc ggacaagggg actcattgca cgaacatatt
     2581 gcgaatcttg ctggttcgcc agccatcaaa aagggcatac tccagacagt caaagtagtg
     2641 gatgagctag ttaaggtcat gggacgtcac aaaccggaaa acattgtaat cgagatggca
     2701 cgcgaaaatc aaacgactca gaaggggcaa aaaaacagtc gagagcggat gaagagaata
     2761 gaagagggta ttaaagaact gggcagccag atcttaaagg agcatcctgt ggaaaatacc
     2821 caattgcaga acgagaaact ttacctctat tacctacaaa atggaaggga catgtatgtt
     2881 gatcaggaac tggacataaa ccgtttatct gattacgacg tcgatgccat tgtaccccaa
     2941 tcctttttga aggacgattc aatcgacaat aaagtgctta cacgctcgga taagaaccga
     3001 gggaaaagtg acaatgttcc aagcgaggaa gtcgtaaaga aaatgaagaa ctattggcgg
     3061 cagctcctaa atgcgaaact gataacgcaa agaaagttcg ataacttaac taaagctgag
     3121 aggggtggct tgtctgaact tgacaaggcc ggatttatta aacgtcagct cgtggaaacc
     3181 cgccaaatca caaagcatgt tgcacagata ctagattccc gaatgaatac gaaatacgac
     3241 gagaacgata agctgattcg ggaagtcaaa gtaatcactt taaagtcaaa attggtgtcg
     3301 gacttcagaa aggattttca attctataaa gttagggaga taaataacta ccaccatgcg
     3361 cacgacgctt atcttaatgc cgtcgtaggg accgcactca ttaagaaata cccgaagcta
     3421 gaaagtgagt ttgtgtatgg tgattacaaa gtttatgacg tccgtaagat gatcgcgaaa
     3481 agcgaacagg agataggcaa ggctacagcc aaatacttct tttattctaa cattatgaat
     3541 ttctttaaga cggaaatcac tctggcaaac ggagagatac gcaaacgacc tttaattgaa
     3601 accaatgggg agacaggtga aatcgtatgg gataagggcc gggacttcgc gacggtgaga
     3661 aaagttttgt ccatgcccca agtcaacata gtaaagaaaa ctgaggtgca gaccggaggg
     3721 ttttcaaagg aatcgattct tccaaaaagg aatagtgata agctcatcgc tcgtaaaaag
     3781 gactgggacc cgaaaaagta cggtggcttc gatagcccta cagttgccta ttctgtccta
     3841 gtagtggcaa aagttgagaa gggaaaatcc aagaaactga agtcagtcaa agaattattg
     3901 gggataacga ttatggagcg ctcgtctttt gaaaagaacc ccatcgactt ccttgaggcg
     3961 aaaggttaca aggaagtaaa aaaggatctc ataattaaac taccaaagta tagtctgttt
     4021 gagttagaaa atggccgaaa acggatgttg gctagcgccg gagagcttca aaaggggaac
     4081 gaactcgcac taccgtctaa atacgtgaat ttcctgtatt tagcgtccca ttacgagaag
     4141 ttgaaaggtt cacctgaaga taacgaacag aagcaacttt ttgttgagca gcacaaacat
     4201 tatctcgacg aaatcataga gcaaatttcg gaattcagta agagagtcat cctagctgat
     4261 gccaatctgg acaaagtatt aagcgcatac aacaagcaca gggataaacc catacgtgag
     4321 caggcggaaa atattatcca tttgtttact cttaccaacc tcggcgctcc agccgcattc
     4381 aagtattttg acacaacgat agatcgcaaa cgatacactt ctaccaagga ggtgctagac
     4441 gcgacactga ttcaccaatc catcacggga ttatatgaaa ctcggataga tttgtcacag
     4501 cttgggggtg acggatcccc caagaagaag aggaaagtct cgagcgacta caaagaccat
     4561 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa ggctgcagga
     4621 ggcggtggaa gcgggcgcgc cgacgcgctg gacgatttcg atctcgacat gctgggttct
     4681 gatgccctcg atgactttga cctggatatg ttgggaagcg acgcattgga tgactttgat
     4741 ctggacatgc tcggctccga tgctctggac gatttcgatc tcgatatgtt ataaccggtc
     4801 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     4861 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4921 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4981 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     5041 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     5101 cgataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5161 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5221 cctagggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5281 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5341 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5401 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5461 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5521 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5581 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5641 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5701 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5761 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5881 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5941 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     6001 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6061 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6121 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6181 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6241 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6301 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6361 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6421 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6481 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6541 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6601 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6661 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6721 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6781 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6841 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6901 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6961 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7021 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7081 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7141 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7201 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7261 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatcgatctc
     7321 ccgatcccct agggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7381 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     7441 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     7501 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
     7561 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     7621 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     7681 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     7741 actatttacg gtaaactgcc cacttggcag tacatcaagt gtatc
//
