LOCUS       Exported               10118 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses CibN-dCas9 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-CibN-dCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10118)
  AUTHORS   Polstein LR, Gersbach CA
  TITLE     A light-inducible CRISPR-Cas9 system for control of endogenous gene 
            activation.
  JOURNAL   Nat Chem Biol. 2015 Feb 9. doi: 10.1038/nchembio.1753.
  PUBMED    25664691
REFERENCE   2  (bases 1 to 10118)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10118
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(66..85)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     276..297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(416..436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        421..750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      601..736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     663..682
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             817..1611
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(871..890)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1481..1500
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1785..1906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1822..1841)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1876..1895
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1955..1971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1955..1971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1968..1990)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1979..1995
                     /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(2003..2033)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2048..2069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2186..2203)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2357..2942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2437..2456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3113..3973)
                     /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     3736..3755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3974..4078)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4153..4172)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4344..4723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4724..4927
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4878..4898
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4972..4991
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4972..4990
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5064..5570
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="NGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSSM
                     ITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKKRKFDTETKDCNE
                     KKKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKEL
                     EKTDYI"
     CDS             5577..5600
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5607..5627
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5637..9740
                     /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             9765..9785
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(9821..9838)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9827..10051
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10097..407
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
       61 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
      121 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
      181 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
      241 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
      301 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
      361 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
      421 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
      481 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      541 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      601 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      661 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
      721 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
      781 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
      841 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
      901 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
      961 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     1021 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     1081 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     1141 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     1201 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     1261 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     1321 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     1381 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     1441 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     1501 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     1561 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     1621 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     1681 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     1741 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     1801 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1861 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     1921 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1981 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2041 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2101 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2161 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2221 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2281 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2341 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2401 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2461 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2521 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2581 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2641 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2701 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2761 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2821 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2881 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2941 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3001 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3061 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3121 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3181 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3241 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3301 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3361 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3421 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3481 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3541 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3601 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3661 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3721 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3781 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3841 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3901 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3961 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4021 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4081 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     4141 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     4201 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     4261 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     4321 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     4381 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     4441 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     4501 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     4561 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     4621 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     4681 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     4741 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     4801 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     4861 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     4921 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     4981 tcactatagg gagacccaag ctggctagcg ccatggacta caaagaccat gacggtgatt
     5041 ataaagatca tgacatcgat ataaatggag ctataggagg tgaccttttg ctcaattttc
     5101 ctgacatgtc ggtcctagag cgccaaaggg ctcacctcaa gtacctcaat cccacctttg
     5161 attctcctct cgccggcttc tttgccgatt cttcaatgat taccggcggc gagatggaca
     5221 gctatctttc gactgccggt ttgaatcttc cgatgatgta cggtgagacg acggtggaag
     5281 gtgattcaag actctcaatt tcgccggaaa cgacgcttgg gactggaaat ttcaagaaac
     5341 ggaagtttga tacagagact aaggattgta atgagaagaa gaagaagatg acgatgaaca
     5401 gagatgacct agtagaagaa ggagaagaag agaagtcgaa aataacagag caaaacaatg
     5461 ggagcacaaa aagcatcaag aagatgaaac acaaagccaa gaaagaagag aacaatttct
     5521 ctaatgattc atctaaagtg acgaaggaat tggagaaaac ggattatatt catcacgatt
     5581 acaaggatga cgatgacaag atggccccca agaagaagag gaaggtgggc cgcggaatgg
     5641 ataagaaata ctcaataggc ttagctatcg gcacaaatag cgtcggatgg gcggtgatca
     5701 ctgatgaata taaggttccg tctaaaaagt tcaaggttct gggaaataca gaccgccaca
     5761 gtatcaaaaa aaatcttata ggggctcttt tatttgacag tggagagaca gcggaagcga
     5821 ctcgtctcaa acggacagct cgtagaaggt atacacgtcg gaagaatcgt atttgttatc
     5881 tacaggagat tttttcaaat gagatggcga aagtagatga tagtttcttt catcgacttg
     5941 aagagtcttt tttggtggaa gaagacaaga agcatgaacg tcatcctatt tttggaaata
     6001 tagtagatga agttgcttat catgagaaat atccaactat ctatcatctg cgaaaaaaat
     6061 tggtagattc tactgataaa gcggatttgc gcttaatcta tttggcctta gcgcatatga
     6121 ttaagtttcg tggtcatttt ttgattgagg gagatttaaa tcctgataat agtgatgtgg
     6181 acaaactatt tatccagttg gtacaaacct acaatcaatt atttgaagaa aaccctatta
     6241 acgcaagtgg agtagatgct aaagcgattc tttctgcacg attgagtaaa tcaagacgat
     6301 tagaaaatct cattgctcag ctccccggtg agaagaaaaa tggcttattt gggaatctca
     6361 ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg gcagaagatg
     6421 ctaaattaca gctttcaaaa gatacttacg atgatgattt agataattta ttggcgcaaa
     6481 ttggagatca atatgctgat ttgtttttgg cagctaagaa tttatcagat gctattttac
     6541 tttcagatat cctaagagta aatactgaaa taactaaggc tcccctatca gcttcaatga
     6601 ttaaacgcta cgatgaacat catcaagact tgactctttt aaaagcttta gttcgacaac
     6661 aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt
     6721 atattgatgg gggagctagc caagaagaat tttataaatt tatcaaacca attttagaaa
     6781 aaatggatgg tactgaggaa ttattggtga aactaaatcg tgaagatttg ctgcgcaagc
     6841 aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag ctgcatgcta
     6901 ttttgagaag acaagaagac ttttatccat ttttaaaaga caatcgtgag aagattgaaa
     6961 aaatcttgac ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc aatagtcgtt
     7021 ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt gaagaagttg
     7081 tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt gataaaaatc
     7141 ttccaaatga aaaagtacta ccaaaacata gtttgcttta tgagtatttt acggtttata
     7201 acgaattgac aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca tttctttcag
     7261 gtgaacagaa gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa gtaaccgtta
     7321 agcaattaaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag
     7381 gagttgaaga tagatttaat gcttcattag gtacctacca tgatttgcta aaaattatta
     7441 aagataaaga ttttttggat aatgaagaaa atgaagatat cttagaggat attgttttaa
     7501 cattgacctt atttgaagat agggagatga ttgaggaaag acttaaaaca tatgctcacc
     7561 tctttgatga taaggtgatg aaacagctta aacgtcgccg ttatactggt tggggacgtt
     7621 tgtctcgaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca atattagatt
     7681 ttttgaaatc agatggtttt gccaatcgca attttatgca gctgatccat gatgatagtt
     7741 tgacatttaa agaagacatt caaaaagcac aagtgtctgg acaaggcgat agtttacatg
     7801 aacatattgc aaatttagct ggtagccctg ctattaaaaa aggtatttta cagactgtaa
     7861 aagttgttga tgaattggtc aaagtaatgg ggcggcataa gccagaaaat atcgttattg
     7921 aaatggcacg tgaaaatcag acaactcaaa agggccagaa aaattcgcga gagcgtatga
     7981 aacgaatcga agaaggtatc aaagaattag gaagtcagat tcttaaagag catcctgttg
     8041 aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat ggaagagaca
     8101 tgtatgtgga ccaagaatta gatattaatc gtttaagtga ttatgatgtc gatgccattg
     8161 ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcttaacg cgttctgata
     8221 aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt agtcaaaaag atgaaaaact
     8281 attggagaca acttctaaac gccaagttaa tcactcaacg taagtttgat aatttaacga
     8341 aagctgaacg tggaggtttg agtgaacttg ataaagctgg ttttatcaaa cgccaattgg
     8401 ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc atgaatacta
     8461 aatacgatga aaatgataaa cttattcgag aggttaaagt gattacctta aaatctaaat
     8521 tagtttctga cttccgaaaa gatttccaat tctataaagt acgtgagatt aacaattacc
     8581 atcatgccca tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt aagaaatatc
     8641 caaaacttga atcggagttt gtctatggtg attataaagt ttatgatgtt cgtaaaatga
     8701 ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa atatttcttt tactctaata
     8761 tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc aaacgccctc
     8821 taatcgaaac taatggggaa actggagaaa ttgtctggga taaagggcga gattttgcca
     8881 cagtgcgcaa agtattgtcc atgccccaag tcaatattgt caagaaaaca gaagtacaga
     8941 caggcggatt ctccaaggag tcaattttac caaaaagaaa ttcggacaag cttattgctc
     9001 gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg gtagcttatt
     9061 cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa tccgttaaag
     9121 agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg attgactttt
     9181 tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta cctaaatata
     9241 gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga gaattacaaa
     9301 aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta gctagtcatt
     9361 atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt gtggagcagc
     9421 ataagcatta tttagatgag attattgagc aaatcagtga attttctaag cgtgttattt
     9481 tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga gacaaaccaa
     9541 tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt ggagctcccg
     9601 ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct acaaaagaag
     9661 ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca cgcattgatt
     9721 tgagtcagct aggaggtgac ccaattgccg gatccaaggc tagcccgaaa aagaaacgca
     9781 aagttgggcg cgcctaactt aagtttaaac cgctgatcag cctcgactgt gccttctagt
     9841 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     9901 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     9961 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
    10021 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
    10081 tctagggggt atccccacgc gccctgtagc ggcgcatt
//
