LOCUS       sequence-310775-       10921 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses dRNase H1-EGFP-dCas9 fusion protein in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10921)
  AUTHORS   Abraham KJ, Khosraviani N, Chan JNY, Gorthi A, Samman A, Zhao DY, 
            Wang M, Bokros M, Vidya E, Ostrowski LA, Oshidari R, Pietrobon V, 
            Patel PS, Algouneh A, Singhania R, Liu Y, Yerlici VT, De Carvalho 
            DD, Ohh M, Dickson BC, Hakem R, Greenblatt JF, Lee S, Bishop AJR, 
            Mekhail K
  TITLE     Nucleolar RNA polymerase II drives ribosome biogenesis.
  JOURNAL   Nature. 2020 Sep;585(7824):298-302. doi: 10.1038/s41586-020-2497-0. 
            Epub 2020 Jul 15.
  PUBMED    32669707
REFERENCE   2  (bases 1 to 10921)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-020-2497-0"; journalName: "Nature"; date: "2020-09";
            volume: "585"; issue: "7824"; pages: "298-302"
COMMENT     Sequence Label: dRED
FEATURES             Location/Qualifiers
     source          1..10921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1731..1751
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1761..1763,1764..1766,1767..2477)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1761..1763,1764..1766,1767..2477)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1806..1827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2067..2086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2414..2435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2526..2546
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2556..2582,2583..2585,2586..5072,5073..5075,
                     5076..6662)
                     /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"
     primer_bind     complement(6695..6712)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6701..6925
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6971..7399
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7058..7077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7268..7289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7408..7428)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7413..7742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7593..7728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7655..7674
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7790..7837
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7856..8230
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8360..8481
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8397..8416)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8451..8470
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8530..8546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8530..8546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8543..8565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8554..8570
                     /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(8578..8584,8585..8602,8603..8608))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8623..8644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8761..8778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8932..9520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9012..9031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9691..10482,10483..10551))
                     /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     10314..10333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10552..10656)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10731..10750)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccact agtccagtgt
      781 ggtggaattc tgcagatatc cagcacagtg gcggccgcgc caccatgagc tggtttctgt
      841 tcctggccca cagagtcgcc ttggccgcct tgccctgccg ccgcggctct cgcaggttcg
      901 ggatgttcta tgccgtgagg aggggccgca agaccggggt ctttctgacc tggaatgagt
      961 gcagagcaca ggtggaccgg tttcctgctg ccagatttaa gaagtttgcc acagaggatg
     1021 aggccagggc ctttgtcagg aaatctgcaa gcccggaagt ttcagaaggg catgaaaatc
     1081 aacatggaca agaatcggag gcgaaagcca gcaagcgact ccgtgagcca ctggatggag
     1141 atggacatga aagcgcagag ccgtatgcaa agcacatgaa gccgagcgtg gagccggcgc
     1201 ctccagttag cagagacacg ttttcctaca tgggagactt cgtcgtcgtc tacactgatg
     1261 gctgctgctc cagtaatggg cgtagaaggc cgcgagcagg aatcggcgtt tactgggggc
     1321 cgggccatcc tttaaatgta ggcattagac ttcctgggcg gcagacaaac caaagagcgg
     1381 aaattcatgc agcctgcaaa gccattgaac aagcaaagac tcaaaacatc aataaactgg
     1441 ttctgtatac agacagtatg tttacgataa atggtataac taactgggtt caaggttgga
     1501 agaaaaatgg gtggaagaca agtgcaggga aagaggtgat caacaaagag gactttgtgg
     1561 cactggagag gcttacccag gggatggaca ttcagtggat gcatgttcct ggtcattcgg
     1621 gatttatagg caatgaagaa gctgacagat tagccagaga aggagctaaa caatcggaag
     1681 acggaggtgg atctggaggt ggatctggag gtggatctgg aggtggatct cccaagaaga
     1741 agaggaaggt gggaggttct atggtgagca agggcgagga gctgttcacc ggggtggtgc
     1801 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1861 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1921 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1981 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2041 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2101 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2161 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2221 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2281 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2341 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2401 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2461 tggacgagct gtacaaggga ggtggatctg gaggtggatc tggaggtgga tctggaggtg
     2521 gatctcccaa gaagaagagg aaggtgggag gttctatgga taagaaatac tcaataggct
     2581 tagctatcgg cacaaatagc gtcggatggg cggtgatcac tgatgaatat aaggttccgt
     2641 ctaaaaagtt caaggttctg ggaaatacag accgccacag tatcaaaaaa aatcttatag
     2701 gggctctttt atttgacagt ggagagacag cggaagcgac tcgtctcaaa cggacagctc
     2761 gtagaaggta tacacgtcgg aagaatcgta tttgttatct acaggagatt ttttcaaatg
     2821 agatggcgaa agtagatgat agtttctttc atcgacttga agagtctttt ttggtggaag
     2881 aagacaagaa gcatgaacgt catcctattt ttggaaatat agtagatgaa gttgcttatc
     2941 atgagaaata tccaactatc tatcatctgc gaaaaaaatt ggtagattct actgataaag
     3001 cggatttgcg cttaatctat ttggccttag cgcatatgat taagtttcgt ggtcattttt
     3061 tgattgaggg agatttaaat cctgataata gtgatgtgga caaactattt atccagttgg
     3121 tacaaaccta caatcaatta tttgaagaaa accctattaa cgcaagtgga gtagatgcta
     3181 aagcgattct ttctgcacga ttgagtaaat caagacgatt agaaaatctc attgctcagc
     3241 tccccggtga gaagaaaaat ggcttatttg ggaatctcat tgctttgtca ttgggtttga
     3301 cccctaattt taaatcaaat tttgatttgg cagaagatgc taaattacag ctttcaaaag
     3361 atacttacga tgatgattta gataatttat tggcgcaaat tggagatcaa tatgctgatt
     3421 tgtttttggc agctaagaat ttatcagatg ctattttact ttcagatatc ctaagagtaa
     3481 atactgaaat aactaaggct cccctatcag cttcaatgat taaacgctac gatgaacatc
     3541 atcaagactt gactctttta aaagctttag ttcgacaaca acttccagaa aagtataaag
     3601 aaatcttttt tgatcaatca aaaaacggat atgcaggtta tattgatggg ggagctagcc
     3661 aagaagaatt ttataaattt atcaaaccaa ttttagaaaa aatggatggt actgaggaat
     3721 tattggtgaa actaaatcgt gaagatttgc tgcgcaagca acggaccttt gacaacggct
     3781 ctattcccca tcaaattcac ttgggtgagc tgcatgctat tttgagaaga caagaagact
     3841 tttatccatt tttaaaagac aatcgtgaga agattgaaaa aatcttgact tttcgaattc
     3901 cttattatgt tggtccattg gcgcgtggca atagtcgttt tgcatggatg actcggaagt
     3961 ctgaagaaac aattacccca tggaattttg aagaagttgt cgataaaggt gcttcagctc
     4021 aatcatttat tgaacgcatg acaaactttg ataaaaatct tccaaatgaa aaagtactac
     4081 caaaacatag tttgctttat gagtatttta cggtttataa cgaattgaca aaggtcaaat
     4141 atgttactga aggaatgcga aaaccagcat ttctttcagg tgaacagaag aaagccattg
     4201 ttgatttact cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa gaagattatt
     4261 tcaaaaaaat agaatgtttt gatagtgttg aaatttcagg agttgaagat agatttaatg
     4321 cttcattagg tacctaccat gatttgctaa aaattattaa agataaagat tttttggata
     4381 atgaagaaaa tgaagatatc ttagaggata ttgttttaac attgacctta tttgaagata
     4441 gggagatgat tgaggaaaga cttaaaacat atgctcacct ctttgatgat aaggtgatga
     4501 aacagcttaa acgtcgccgt tatactggtt ggggacgttt gtctcgaaaa ttgattaatg
     4561 gtattaggga taagcaatct ggcaaaacaa tattagattt tttgaaatca gatggttttg
     4621 ccaatcgcaa ttttatgcag ctgatccatg atgatagttt gacatttaaa gaagacattc
     4681 aaaaagcaca agtgtctgga caaggcgata gtttacatga acatattgca aatttagctg
     4741 gtagccctgc tattaaaaaa ggtattttac agactgtaaa agttgttgat gaattggtca
     4801 aagtaatggg gcggcataag ccagaaaata tcgttattga aatggcacgt gaaaatcaga
     4861 caactcaaaa gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa gaaggtatca
     4921 aagaattagg aagtcagatt cttaaagagc atcctgttga aaatactcaa ttgcaaaatg
     4981 aaaagctcta tctctattat ctccaaaatg gaagagacat gtatgtggac caagaattag
     5041 atattaatcg tttaagtgat tatgatgtcg atgccattgt tccacaaagt ttccttaaag
     5101 acgattcaat agacaataag gtcttaacgc gttctgataa aaatcgtggt aaatcggata
     5161 acgttccaag tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa cttctaaacg
     5221 ccaagttaat cactcaacgt aagtttgata atttaacgaa agctgaacgt ggaggtttga
     5281 gtgaacttga taaagctggt tttatcaaac gccaattggt tgaaactcgc caaatcacta
     5341 agcatgtggc acaaattttg gatagtcgca tgaatactaa atacgatgaa aatgataaac
     5401 ttattcgaga ggttaaagtg attaccttaa aatctaaatt agtttctgac ttccgaaaag
     5461 atttccaatt ctataaagta cgtgagatta acaattacca tcatgcccat gatgcgtatc
     5521 taaatgccgt cgttggaact gctttgatta agaaatatcc aaaacttgaa tcggagtttg
     5581 tctatggtga ttataaagtt tatgatgttc gtaaaatgat tgctaagtct gagcaagaaa
     5641 taggcaaagc aaccgcaaaa tatttctttt actctaatat catgaacttc ttcaaaacag
     5701 aaattacact tgcaaatgga gagattcgca aacgccctct aatcgaaact aatggggaaa
     5761 ctggagaaat tgtctgggat aaagggcgag attttgccac agtgcgcaaa gtattgtcca
     5821 tgccccaagt caatattgtc aagaaaacag aagtacagac aggcggattc tccaaggagt
     5881 caattttacc aaaaagaaat tcggacaagc ttattgctcg taaaaaagac tgggatccaa
     5941 aaaaatatgg tggttttgat agtccaacgg tagcttattc agtcctagtg gttgctaagg
     6001 tggaaaaagg gaaatcgaag aagttaaaat ccgttaaaga gttactaggg atcacaatta
     6061 tggaaagaag ttcctttgaa aaaaatccga ttgacttttt agaagctaaa ggatataagg
     6121 aagttaaaaa agacttaatc attaaactac ctaaatatag tctttttgag ttagaaaacg
     6181 gtcgtaaacg gatgctggct agtgccggag aattacaaaa aggaaatgag ctggctctgc
     6241 caagcaaata tgtgaatttt ttatatttag ctagtcatta tgaaaagttg aagggtagtc
     6301 cagaagataa cgaacaaaaa caattgtttg tggagcagca taagcattat ttagatgaga
     6361 ttattgagca aatcagtgaa ttttctaagc gtgttatttt agcagatgcc aatttagata
     6421 aagttcttag tgcatataac aaacatagag acaaaccaat acgtgaacaa gcagaaaata
     6481 ttattcattt atttacgttg acgaatcttg gagctcccgc tgcttttaaa tattttgata
     6541 caacaattga tcgtaaacga tatacgtcta caaaagaagt tttagatgcc actcttatcc
     6601 atcaatccat cactggtctt tatgaaacac gcattgattt gagtcagcta ggaggtgact
     6661 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     6721 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     6781 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     6841 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     6901 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     6961 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7021 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7081 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7141 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     7201 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7261 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7321 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7381 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     7441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     7501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     7561 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     7621 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7681 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     7741 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     7801 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     7861 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     7921 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     7981 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     8041 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     8101 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     8161 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     8221 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     8281 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     8341 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     8401 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     8461 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     8521 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     8581 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     8641 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     8701 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     8761 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     8821 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     8881 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     8941 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     9001 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     9061 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     9121 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     9181 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     9241 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     9301 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     9361 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     9421 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     9481 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     9541 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     9601 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     9661 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     9721 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     9781 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     9841 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     9901 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     9961 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    10021 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    10081 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    10141 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    10201 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    10261 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    10321 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    10381 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    10441 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    10501 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    10561 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    10621 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    10681 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
    10741 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
    10801 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
    10861 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
    10921 t
//
