CRISPRoff-v2.1

LOCUS       CRISPRoff-v2.1               11885 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses CRISPRoff-v2.1 
            (DNMT3A-DNMT3L-XTEN80-dCas9-HA-2xNLS-BFP-KRAB) downstream of the CAG
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    CRISPRoff-v2.1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11885)
  AUTHORS   Nunez JK, Chen J, Pommier GC, Cogan JZ, Replogle JM, Adriaens C, 
            Ramadoss GN, Shi Q, Hung KL, Samelson AJ, Pogson AN, Kim JYS, Chung 
            A, Leonetti MD, Chang HY, Kampmann M, Bernstein BE, Hovestadt V, 
            Gilbert LA, Weissman JS
  TITLE     Genome-wide programmable transcriptional memory by CRISPR-based 
            epigenome editing.
  JOURNAL   Cell. 2021 Apr 7. pii: S0092-8674(21)00353-6. doi: 
            10.1016/j.cell.2021.03.025.
  PUBMED    33838111
REFERENCE   2  (bases 1 to 11885)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11885
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3650..7753
                     /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             7757..7783
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7802..7822
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7829..7849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7889..8584
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             8630..8815
                     /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"
     primer_bind     complement(8907..8926)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8972..9027
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9026..9045)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(9388..9404)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9388..9404)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9401..9423)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9412..9428
                     /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(9436..9466)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9481..9502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9561..9756
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9607..9742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9669..9688
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    9762..9896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9799..9818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9853..9872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9964..9981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10135..10723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10215..10234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(10894..11754)
                     /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     11517..11536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11755..11859)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggncgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttctgcagtc
     1741 gacggtaccg cgggcccggg atccaccggt cgccaccatg aaccatgacc aggaatttga
     1801 ccccccaaag gtttacccac ctgtgccagc tgagaagagg aagcccatcc gcgtgctgtc
     1861 tctctttgat gggattgcta cagggctcct ggtgctgaag gacctgggca tccaagtgga
     1921 ccgctacatt gcctccgagg tgtgtgagga ctccatcacg gtgggcatgg tgcggcacca
     1981 gggaaagatc atgtacgtcg gggacgtccg cagcgtcaca cagaagcata tccaggagtg
     2041 gggcccattc gacctggtga ttggaggcag tccctgcaat gacctctcca ttgtcaaccc
     2101 tgcccgcaag ggactttatg agggtactgg ccgcctcttc tttgagttct accgcctcct
     2161 gcatgatgcg cggcccaagg agggagatga tcgccccttc ttctggctct ttgagaatgt
     2221 ggtggccatg ggcgttagtg acaagaggga catctcgcga tttcttgagt ctaaccccgt
     2281 gatgattgac gccaaagaag tgtctgctgc acacagggcc cgttacttct ggggtaacct
     2341 tcctggcatg aacaggcctt tggcatccac tgtgaatgat aagctggagc tgcaagagtg
     2401 tctggagcac ggcagaatag ccaagttcag caaagtgagg accattacca ccaggtcaaa
     2461 ctctataaag cagggcaaag accagcattt ccccgtcttc atgaacgaga aggaggacat
     2521 cctgtggtgc actgaaatgg aaagggtgtt tggcttcccc gtccactaca cagacgtctc
     2581 caacatgagc cgcttggcga ggcagagact gctgggccga tcgtggagcg tgccggtcat
     2641 ccgccacctc ttcgctccgc tgaaggaata ttttgcttgt gtgtctagcg gcaatagtaa
     2701 cgctaacagc cgcgggccga gcttcagcag cggcctggtg ccgttaagct tgcgcggcag
     2761 ccatatgggc cctatggaga tatacaagac agtgtctgca tggaagagac agccagtgcg
     2821 ggtactgagc ctcttcagaa acatcgacaa ggtactaaag agtttgggct tcttggaaag
     2881 cggttctggt tctgggggag gaacgctgaa gtacgtggaa gatgtcacaa atgtcgtgag
     2941 gagagacgtg gagaaatggg gcccctttga cctggtgtac ggctcgacgc agcccctagg
     3001 cagctcttgt gatcgctgtc ccggctggta catgttccag ttccaccgga tcctgcagta
     3061 tgcgctgcct cgccaggaga gtcagcggcc cttcttctgg atattcatgg acaatctgct
     3121 gctgactgag gatgaccaag agacaactac ccgcttcctt cagacagagg ctgtgaccct
     3181 ccaggatgtc cgtggcagag actaccagaa tgctatgcgg gtgtggagca acattccagg
     3241 gctgaagagc aagcatgcgc ccctgacccc aaaggaagaa gagtatctgc aagcccaagt
     3301 cagaagcagg agcaagctgg acgccccgaa agttgacctc ctggtgaaga actgccttct
     3361 cccgctgaga gagtacttca agtatttttc tcaaaactca cttcctcttg gagggccgag
     3421 ctctggcgca cccccaccaa gtggagggtc tcctgccggg tccccaacat ctactgaaga
     3481 aggcaccagc gaatccgcaa cgcccgagtc aggccctggt acctccacag aaccatctga
     3541 aggtagtgcg cctggttccc cagctggaag ccctacttcc accgaagaag gcacgtcaac
     3601 cgaaccaagt gaaggatctg cccctgggac cagcactgaa ccatctgaga tggacaagaa
     3661 gtattctatc ggactggcca tcgggactaa tagcgtcggg tgggccgtga tcactgacga
     3721 gtacaaggtg ccctctaaga agttcaaggt gctcgggaac accgaccggc attccatcaa
     3781 gaaaaatctg atcggagctc tcctctttga ttcaggggag accgctgaag caacccgcct
     3841 caagcggact gctagacggc ggtacaccag gaggaagaac cggatttgtt accttcaaga
     3901 gatattctcc aacgaaatgg caaaggtcga cgacagcttc ttccataggc tggaagaatc
     3961 attcctcgtg gaagaggata agaagcatga acggcatccc atcttcggta atatcgtcga
     4021 cgaggtggcc tatcacgaga aatacccaac catctaccat cttcgcaaaa agctggtgga
     4081 ctcaaccgac aaggcagacc tccggcttat ctacctggcc ctggcccaca tgatcaagtt
     4141 cagaggccac ttcctgatcg agggcgacct caatcctgac aatagcgatg tggataaact
     4201 gttcatccag ctggtgcaga cttacaacca gctctttgaa gagaacccca tcaatgcaag
     4261 cggagtcgat gccaaggcca ttctgtcagc ccggctgtca aagagccgca gacttgagaa
     4321 tcttatcgct cagctgccgg gtgaaaagaa aaatggactg ttcgggaacc tgattgctct
     4381 ttcacttggg ctgactccca atttcaagtc taatttcgac ctggcagagg atgccaagct
     4441 gcaactgtcc aaggacacct atgatgacga tctcgacaac ctcctggccc agatcggtga
     4501 ccaatacgcc gaccttttcc ttgctgctaa gaatctttct gacgccatcc tgctgtctga
     4561 cattctccgc gtgaacactg aaatcaccaa ggcccctctt tcagcttcaa tgattaagcg
     4621 gtatgatgag caccaccagg acctgaccct gcttaaggca ctcgtccggc agcagcttcc
     4681 ggagaagtac aaggaaatct tctttgacca gtcaaagaat ggatacgccg gctacatcga
     4741 cggaggtgcc tcccaagagg aattttataa gtttatcaaa cctatccttg agaagatgga
     4801 cggcaccgaa gagctcctcg tgaaactgaa tcgggaggat ctgctgcgga agcagcgcac
     4861 tttcgacaat gggagcattc cccaccagat ccatcttggg gagcttcacg ccatccttcg
     4921 gcgccaagag gacttctacc cctttcttaa ggacaacagg gagaagattg agaaaattct
     4981 cactttccgc atcccctact acgtgggacc cctcgccaga ggaaatagcc ggtttgcttg
     5041 gatgaccaga aagtcagaag aaactatcac tccctggaac ttcgaagagg tggtggacaa
     5101 gggagccagc gctcagtcat tcatcgaacg gatgactaac ttcgataaga acctccccaa
     5161 tgagaaggtc ctgccgaaac attccctgct ctacgagtac tttaccgtgt acaacgagct
     5221 gaccaaggtg aaatatgtca ccgaagggat gaggaagccc gcattcctgt caggcgaaca
     5281 aaagaaggca attgtggacc ttctgttcaa gaccaataga aaggtgaccg tgaagcagct
     5341 gaaggaggac tatttcaaga aaattgaatg cttcgactct gtggagatta gcggggtcga
     5401 agatcggttc aacgcaagcc tgggtaccta ccatgatctg cttaagatca tcaaggacaa
     5461 ggattttctg gacaatgagg agaacgagga catccttgag gacattgtcc tgactctcac
     5521 tctgttcgag gaccgggaaa tgatcgagga gaggcttaag acctacgccc atctgttcga
     5581 cgataaagtg atgaagcaac ttaaacggag aagatatacc ggatggggac gccttagccg
     5641 caaactcatc aacggaatcc gggacaaaca gagcggaaag accattcttg atttccttaa
     5701 gagcgacgga ttcgctaatc gcaacttcat gcaacttatc catgatgatt ccctgacctt
     5761 taaggaggac atccagaagg cccaagtgtc tggacaaggt gactcactgc acgagcatat
     5821 cgcaaatctg gctggttcac ccgctattaa gaagggtatt ctccagaccg tgaaagtcgt
     5881 ggacgagctg gtcaaggtga tgggtcgcca taaaccagag aacattgtca tcgagatggc
     5941 cagggaaaac cagactaccc agaagggaca gaagaacagc agggagcgga tgaaaagaat
     6001 tgaggaaggg attaaggagc tcgggtcaca gatccttaaa gagcacccgg tggaaaacac
     6061 ccagcttcag aatgagaagc tctatctgta ctaccttcaa aatggacgcg atatgtatgt
     6121 ggaccaagag cttgatatca acaggctctc agactacgac gtggacgcca tcgtccctca
     6181 gagcttcctc aaagacgact caattgacaa taaggtgctg actcgctcag acaagaaccg
     6241 gggaaagtca gataacgtgc cctcagagga agtcgtgaaa aagatgaaga actattggcg
     6301 ccagcttctg aacgcaaagc tgatcactca gcggaagttc gacaatctca ctaaggctga
     6361 gaggggcgga ctgagcgaac tggacaaagc aggattcatt aaacggcaac ttgtggagac
     6421 tcggcagatt actaaacatg tcgcccaaat ccttgactca cgcatgaata ccaagtacga
     6481 cgaaaacgac aaacttatcc gcgaggtgaa ggtgattacc ctgaagtcca agctggtcag
     6541 cgatttcaga aaggactttc aattctacaa agtgcgggag atcaataact atcatcatgc
     6601 tcatgacgca tatctgaatg ccgtggtggg aaccgccctg atcaagaagt acccaaagct
     6661 ggaaagcgag ttcgtgtacg gagactacaa ggtctacgac gtgcgcaaga tgattgccaa
     6721 atctgagcag gagatcggaa aggccaccgc aaagtacttc ttctacagca acatcatgaa
     6781 tttcttcaag accgaaatca cccttgcaaa cggtgagatc cggaagaggc cgctcatcga
     6841 gactaatggg gagactggcg aaatcgtgtg ggacaagggc agagatttcg ctaccgtgcg
     6901 caaagtgctt tctatgcctc aagtgaacat cgtgaagaaa accgaggtgc aaaccggagg
     6961 cttttctaag gaatcaatcc tccccaagcg caactccgac aagctcattg caaggaagaa
     7021 ggattgggac cctaagaagt acggcggatt cgattcacca actgtggctt attctgtcct
     7081 ggtcgtggct aaggtggaaa aaggaaagtc taagaagctc aagagcgtga aggaactgct
     7141 gggtatcacc attatggagc gcagctcctt cgagaagaac ccaattgact ttctcgaagc
     7201 caaaggttac aaggaagtca agaaggacct tatcatcaag ctcccaaagt atagcctgtt
     7261 cgaactggag aatgggcgga agcggatgct cgcctccgct ggcgaacttc agaagggtaa
     7321 tgagctggct ctcccctcca agtacgtgaa tttcctctac cttgcaagcc attacgagaa
     7381 gctgaagggg agccccgagg acaacgagca aaagcaactg tttgtggagc agcataagca
     7441 ttatctggac gagatcattg agcagatttc cgagttttct aaacgcgtca ttctcgctga
     7501 tgccaacctc gataaagtcc ttagcgcata caataagcac agagacaaac caattcggga
     7561 gcaggctgag aatatcatcc acctgttcac cctcaccaat cttggtgccc ctgccgcatt
     7621 caagtacttc gacaccacca tcgaccggaa acgctatacc tccaccaaag aagtgctgga
     7681 cgccaccctc atccaccaga gcatcaccgg actttacgaa actcggattg acctctcaca
     7741 gctcggaggg gatgcctatc cctatgacgt gcccgattat gccagcctgg gcagcggctc
     7801 ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag cggaaagtgg acggcattgg
     7861 tagtgggagc aacggcagca gcggatccag cgagctgatt aaggagaaca tgcacatgaa
     7921 gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg
     7981 caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc
     8041 cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca
     8101 cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca catgggagag
     8161 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
     8221 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt
     8281 gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg
     8341 cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc
     8401 aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt
     8461 ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga
     8521 gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct
     8581 taatggcggt ggcggaggga tggatgctaa gtcactaact gcctggtccc ggacactggt
     8641 gaccttcaag gatgtatttg tggacttcac cagggaggag tggaagctgc tggacactgc
     8701 tcagcagatc gtgtacagaa atgtgatgct ggagaactat aagaacctgg tttccttggg
     8761 ttatcagctt actaagccag atgtgatcct ccggttggag aagggagaag agccctagta
     8821 agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
     8881 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
     8941 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
     9001 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
     9061 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
     9121 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
     9181 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
     9241 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
     9301 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
     9361 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     9421 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     9481 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     9541 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     9601 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     9661 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     9721 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     9781 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     9841 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     9901 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     9961 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
    10021 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
    10081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
    10141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
    10201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
    10261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
    10321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
    10381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
    10441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
    10501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
    10561 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
    10621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
    10681 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
    10741 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
    10801 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
    10861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
    10921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
    10981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
    11041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
    11101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
    11161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
    11221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
    11281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
    11341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
    11401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
    11461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
    11521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
    11581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
    11641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
    11701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
    11761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
    11821 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    11881 acctg
//