pdCas9-Tet1-CD

LOCUS       pdCas9-Tet1-CD               10468 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  To introduce dCas9-fused Tet1-CD and sgRNA2.0 system.
ACCESSION   .
VERSION     .
KEYWORDS    pdCas9-Tet1-CD
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10468)
  AUTHORS   Xu X, Tao Y, Gao X, Zhang L, Li X, Zou W, Ruan K, Wang F, Xu GL, Hu 
            R
  TITLE     A CRISPR-based approach for targeted DNA demethylation.
  JOURNAL   Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9. 
            eCollection 2016.
  PUBMED    27462456
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|pdCas9-Tet1-CD|
FEATURES             Location/Qualifiers
     source          1..10468
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        285..303
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        355..373
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     enhancer        416..701
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        703..980
                     /label=chicken beta-actin promoter
     intron          981..1209
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1230..1256
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1260..1280
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1290..5390
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5394..5414
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7509..7526)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7515..7722
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7731..7871
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7731..7860
                     /label=AAV2 ITR
     rep_origin      7946..8401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8033..8052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8243..8264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8418..8437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8537..8559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8597..8615)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8683..8787
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8788..9648
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9006..9025)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9819..10407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10308..10327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag gccaacatga ggatcaccca
      301 tgtctgcagg gcctagcaag ttaaaataag gctagtccgt tatcaacttg gccaacatga
      361 ggatcaccca tgtctgcagg gccaagtggc accgagtcgg tgcttttttg gtacccgtta
      421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      481 caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      541 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      601 atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta
      661 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
      721 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
      781 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg
      841 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
      901 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      961 agcgaagcgc gcggcgggcg ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc
     1021 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
     1081 gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag
     1141 ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact
     1201 ttttttcagg ttggaccggt gccaccatgt acccatacga tgttccagat tacgcttcgc
     1261 cgaagaaaaa gcgcaaggtc gaagcgtccg acaagaagta cagcatcggc ctggccatcg
     1321 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     1381 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     1441 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     1501 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     1561 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     1621 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     1681 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     1741 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     1801 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     1861 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     1921 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     1981 agaagaagaa tggcctgttc ggcaacctga ttgccctgag cctgggcctg acccccaact
     2041 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     2101 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     2161 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     2221 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     2281 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     2341 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     2401 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     2461 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     2521 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     2581 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     2641 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     2701 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     2761 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     2821 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     2881 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     2941 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     3001 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     3061 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     3121 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     3181 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     3241 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     3301 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     3361 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     3421 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     3481 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     3541 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     3601 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     3661 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     3721 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     3781 ggctgtccga ctacgatgtg gacgctatcg tgcctcagag ctttctgaag gacgactcca
     3841 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     3901 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     3961 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     4021 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     4081 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     4141 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     4201 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     4261 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     4321 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     4381 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     4441 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     4501 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     4561 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     4621 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     4681 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     4741 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     4801 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     4861 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     4921 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     4981 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     5041 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     5101 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     5161 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     5221 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     5281 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     5341 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac agccccaaga
     5401 agaagagaaa ggtggaggcc agcggtggag gaagtggagg tggttcaggt ggaggatccg
     5461 gcgaagctgc accctgtgac tgtgatggag gtacacaaaa agaaaaaggc ccatattata
     5521 cacaccttgg ggcaggacca agtgtggctg ctgtcaggga gctcatggag actaggtttg
     5581 gccagaaggg gaaggcaatc cggattgaga agatagtgtt cacggggaag gaagggaaga
     5641 gctctcaggg ctgcccggtc gccaagtggg tgatcagaag aagtggtcct gaagagaagc
     5701 ttatttgttt ggttcgtgag cgtgtagacc atcactgttc gacggctgtg atagttgtcc
     5761 ttatcctgct gtgggaaggt atccctcgcc tgatggctga ccgcctgtac aaagagctca
     5821 ctgagaactt gaggtcctac agcggacatc ccacagaccg aagatgtacc ctcaacaaaa
     5881 agcgtacctg cacctgtcaa ggcatcgacc caaaaacctg cggagcgtcc ttctcctttg
     5941 gctgttcgtg gagcatgtat ttcaacggct gtaagtttgg gaggagtgaa aaccccagaa
     6001 aattcagact tgctccaaac taccccttac ataactacta taagagaatt actggaatga
     6061 gttctgaagg aagtgacgtg aaaaccgggt ggatcattcc agaccgcaag accctcataa
     6121 gcagagagga aaaacagctt gaaaagaatt tacaagaatt ggctacagta ttagctccac
     6181 tttacaagca gatggctcca gttgcttatc aaaatcaggt ggaatatgaa gaagttgctg
     6241 gagactgtcg acttggaaat gaagaggggc gtcctttctc tggtgtcacc tgttgcatgg
     6301 atttttgtgc ccattctcac aaggacattc acaacatgca caacggaagc accgtggtgt
     6361 gtacgttgat tcgagcagat ggccgtgaca caaattgtcc cgaggatgaa caactccacg
     6421 tcctgccact ataccggctt gcagacactg atgaatttgg ctccgtggaa gggatgaagg
     6481 ccaaaatcaa atctggggcc atccaagtca atgggccaac caggaagagg cgactacgtt
     6541 ttactgagcc tgttcctcga tgtgggaaga gggccaaaat gaagcagaac cacaataaat
     6601 caggttcaca caacactaag agcttttcat cagcctcatc tacttctcac ctagtgaaag
     6661 acgaatctac agacttctgt cccctgcagg cttcctccgc agaaacatct acctgtacgt
     6721 acagtaaaac agcctcaggt gggtttgcag aaacaagtag tattctccac tgcacaatgc
     6781 cttctggagc acacagtggt gctaatgcag ctgctgggga atgtactgga acggtgcagc
     6841 ctgccgaggt ggctgctcat cctcaccagt ctcttcccac agccgattct cccgttcatg
     6901 ctgagcctct cactagtcca tctgagcagc taacttctaa ccagtcaaac cagcagctcc
     6961 ctctcctcag caattctcag aaactggctt cctgtcaggt ggaagatgag cggcaccctg
     7021 aagcggatga gcctcagcac cccgaggacg ataacttgcc tcaacttgat gagttctggt
     7081 cagacagtga ggagatctac gccgatcctt cctttggtgg cgtggcgata gcacccattc
     7141 acggctcggt gctcattgag tgcgctcgga aggagcttca tgctaccacc tctttgcgct
     7201 cccccaaacg aggggtccct tttcgtgtgt cccttgtatt ctaccagcac aaaagcctaa
     7261 acaagcctaa tcatggtttt gatatcaaca aaattaagtg taaatgcaaa aaagtaacga
     7321 aaaaaaagcc cgcagaccgg gagtgtcctg atgtatcccc cgaagccaat ttatcacacc
     7381 aaattccttc tcgagttgca tcaaccttaa cccgagacaa tgttgttacc gtgtccccat
     7441 actctctcac tcatgttgcg ggaccctaca atcgttgggt ctaagaattc ctagagctcg
     7501 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     7561 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     7621 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     7681 caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct
     7741 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     7801 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     7861 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     7921 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     7981 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     8041 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     8101 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     8161 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     8221 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     8281 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     8341 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     8401 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     8461 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     8521 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     8581 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     8641 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     8701 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     8761 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     8821 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     8881 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     8941 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     9001 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     9061 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     9121 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     9181 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     9241 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     9301 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     9361 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     9421 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     9481 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
     9541 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     9601 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     9661 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     9721 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     9781 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     9841 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     9901 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     9961 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
    10021 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
    10081 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
    10141 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
    10201 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
    10261 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
    10321 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
    10381 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
    10441 gccttttgct ggccttttgc tcacatgt
//