LOCUS       Exported               15637 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of Candida albicans codon-optimized Cas9 (CaCas9) - 
            integrates at ENO1 (Part of Duet system).
ACCESSION   .
VERSION     .
KEYWORDS    pV1025
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15637)
  AUTHORS   Vyas VK, Barrasa MI, Fink GR
  TITLE     A CRISPR system permits genetic engineering of essential genes and 
            gene families.
  JOURNAL   Sci Adv. 2015;1(3):e1500248.
  PUBMED    25977940
REFERENCE   2  (bases 1 to 15637)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15637
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(19..474)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(156..177)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     368..387
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     601..623
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     615..632
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     616..632
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     642..661
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        642..660
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2687..6790
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6803..6823
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6827..6847
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6851..6871
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6872..6937
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      7002..7249
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7018..7036)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(7268..7301)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(7303..7319)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8398..9669
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     terminator      10096..10293
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(10894..11237)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    complement(11278..11311)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(11313..11329)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13465..13485)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(13465..13483)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13504..13520)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13504..13520)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13517..13539)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13528..13544
                     /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(13552..13582)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13597..13618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13735..13752)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13906..14494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13986..14005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14665..15525)
                     /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     15288..15307
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15526..15630)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
       61 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
      121 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
      181 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
      241 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
      301 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
      361 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
      421 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
      481 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
      541 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
      601 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
      661 gcgaattgag gcctgcatgc ggtaccttta tcagcaagta gaaaacaacc aaagctcttg
      721 aaattgtgca atgaagattt catcaaacat aaaaccatta tgaacgcttg gaagttgtta
      781 caacgaagaa gaataaccca acaatctgaa aaattgtcta agcaatataa aagcattgtc
      841 aatgccatgg aagatttgaa gcaaacaagt cccgaattgt tcgaagctgc aaatgctaaa
      901 gaccctaaac gtttcactac cttcccaata gagatgagag tgcctaccga ttatccacct
      961 aacaagccat ggacttacaa ctttgtttct tcaaaaaccc atcattagac tgggttcaga
     1021 tgtaaataga tattatatta taaatgtaca taatcgaata gattgttatt atttgttcaa
     1081 ctcgtcctaa tcctccaata ctctcgcctt tctttttcta ctaggtgtgc cactactacc
     1141 actgggcgtc tcgttcttct cgatactatt agctttactt cctgcactag cagtggttgg
     1201 atcaacagaa tcttcataat catcaaaatc gtcttttgaa gaccccccgt ttgatgtatg
     1261 gccctgtctt ttcatcaaac tttttatata gttgactgaa ctgaggctaa atatgtgatc
     1321 atcttcacta tagacaatct ttctcttatt tgcaccaccg ccaccaccag tctttgagaa
     1381 attctcaaaa ccttttacga tattaccaag cgggctctct tcgaaataat ctatctcttt
     1441 ttgatatatc gaatcctcta gcgtggttag ctttctagtt agttcttgct tcttaagaat
     1501 ttgctggatt agtttatttt tcaattcaac gtatttctca gagtcatctt tagattttga
     1561 tgaagatgtg cgttcattcg ctatatcctt cttggtcgtg tcttttcgat cctccttggc
     1621 tggcactgaa ctcgtctttt ttggcgttgc tgttccagac agacttatct cattagattt
     1681 ggaacttgtg ggtttaacat catttgcatc tttagtagac atgattgtgt aataccgtga
     1741 ttgtttgttt tgaaaggtct gtcatatttc tatcaatttc aaaacaaaat gttcatcaga
     1801 aaaaagccaa aaatgtctct tctagcttct tagtggtgtc gcaaaataca caatgtcgct
     1861 caacaatcca cattcccggc gcatagctca aatcacatga ctacagctaa caattacaca
     1921 aaaaaattct cttttcgatg tagcaactat cttcaactaa aacattttct ccttcggccc
     1981 atgattgtcc tccgggtcga cagcaagccg ttacaattga gatggaaagc gacctacctt
     2041 cactcgataa ggtgcttaat tgtacttcat ataaatctgg cccggatcta aacaaatgag
     2101 ttccattaag ccgtgggttc tcaattaggg tttttgtttt tgatttagaa aaaagagatc
     2161 aagatttgtt tacaggtgat gccttttttt agaacttatg cgttgcaaaa gttgactaac
     2221 gatttctata aggtgatcca cactaattat acaaacgtac aaacagacat acttttcctg
     2281 cgttcacctg atgttggcca gatttctctc ttcattgcat agaacataac cacactaggg
     2341 caacagaaaa aaaaaaaaaa agtgcatcgg gaagttgtgt tccattcatt atatgtctac
     2401 tactgcatat gagtagccca cccaccacca ccatagtaag tttttgtgta tgcgcgccgt
     2461 caggttattt catttctgaa tttttcaacc accttactcc ctttattgtt gattgacaat
     2521 tttgctcaca gtaagatctt ttagactcca attaatataa aataagtctg attttccaat
     2581 tcctgttttt tctttttttt tctgtttcta tttctttcct tttctccctt ttttttaatt
     2641 cttcattcaa tcatcaattg ataattcagg aatattacaa cccgggatgg ataaaaagta
     2701 tagtattggt ttagatattg gtactaactc tgtgggttgg gcagttatca ctgacgaata
     2761 taaagttcca tcaaagaaat ttaaggtgtt aggtaacact gacagacact caataaaaaa
     2821 gaatcttatc ggtgctcttt tgttcgactc cggtgaaact gccgaggcta cacgtttaaa
     2881 aagaacagca agaagaagat atacccgtag aaaaaataga atatgttatt tacaagaaat
     2941 cttttctaat gaaatggcta aagttgatga ttcctttttc catagattgg aagagtcatt
     3001 tttggttgaa gaagacaaaa agcatgagag acatccaatc tttgggaata tagttgatga
     3061 agtggcttac catgaaaaat atcctaccat ttatcattta agaaagaaat tggtagattc
     3121 aactgataaa gctgacctta gattaatcta tttagcactt gcccatatga ttaaatttag
     3181 aggtcatttt ttgattgaag gtgatttgaa cccagataat tctgacgtgg ataaattatt
     3241 tattcaatta gtccaaacct acaaccaatt atttgaggaa aatccaatta atgctagtgg
     3301 tgtcgatgcc aaagctatat tatcagccag attatcaaaa tctagacgtt tggaaaattt
     3361 gattgcccaa ttgccaggag aaaaaaagaa tggattattt ggaaacttga tcgcattatc
     3421 attgggtttg acaccaaatt ttaaatctaa ttttgattta gctgaagatg ctaaattaca
     3481 attatcaaaa gacacctatg acgacgattt ggacaattta cttgctcaaa ttggtgatca
     3541 atatgcagat ttgttcttag ctgctaaaaa cttatctgat gctattttgt tgtctgatat
     3601 tttgagagtg aacacagaaa taaccaaagc tccattatca gcatctatga tcaaacgtta
     3661 tgatgaacac catcaggatt tgactttatt gaaagctttg gtgagacaac aattgccaga
     3721 gaagtataaa gaaatctttt tcgatcaatc taaaaacggg tatgcaggtt atattgatgg
     3781 gggtgcctcc caagaggaat tttacaaatt tataaaacct attttagaaa agatggatgg
     3841 gactgaggaa cttttggtca aattgaacag agaagatttg ttacgtaaac agagaacttt
     3901 tgataatggt agtatacctc accaaattca tttgggtgag ttgcatgcaa ttttaagaag
     3961 acaagaagat ttttatccat ttttaaaaga taatagagaa aaaatcgaga aaattttaac
     4021 ctttagaatt ccatactatg ttgggccttt ggctagaggt aattcaagat ttgcctggat
     4081 gacacgtaaa tcagaagaaa ctattacccc ttggaatttt gaagaggttg ttgataaagg
     4141 agcatcagca cagagtttta ttgaaagaat gaccaatttc gataaaaact taccaaatga
     4201 aaaagtttta ccaaaacatt ccttgttata cgaatatttt actgtttaca atgaacttac
     4261 aaaggttaaa tatgttactg aaggtatgcg taagccagcc tttttatctg gagaacagaa
     4321 aaaggcaata gttgatttat tgtttaaaac aaatagaaaa gttactgtta aacaattaaa
     4381 agaagattac tttaagaaaa ttgaatgttt tgattcagtt gaaatcagtg gtgttgaaga
     4441 cagatttaat gctagtttag gaacttacca tgatttactt aaaattatca aagataaaga
     4501 tttcttggat aacgaagaaa atgaagacat tttagaagac attgttttaa ccttaacttt
     4561 attcgaagat agagagatga ttgaagaacg tttgaagact tatgcacatt tgtttgacga
     4621 taaagtgatg aaacagttga aaagaagacg ttatactgga tggggtagat tgtctcgtaa
     4681 attgatcaat ggaattagag ataaacaaag tggtaaaact atcttggact ttttgaaatc
     4741 tgacggattt gctaatagaa atttcatgca attgatccac gacgatagtt tgacatttaa
     4801 agaagacatc caaaaggccc aagtgagtgg gcaaggtgat tcattacatg aacatattgc
     4861 aaatttagcc ggatctcctg ctattaagaa agggatatta caaactgtta aagttgtgga
     4921 tgaattagtg aaagtaatgg gaagacataa acctgaaaac attgtcattg agatggcaag
     4981 agaaaatcaa actacacaaa aaggacagaa aaatagtaga gaacgtatga aaagaataga
     5041 agagggtatt aaagaattgg gtagtcaaat attgaaagaa cacccagtgg aaaataccca
     5101 gttgcaaaat gaaaaattat atctttacta ccttcaaaat ggacgtgata tgtatgttga
     5161 tcaggaatta gatataaata gactttcaga ttatgatgta gatcatatag ttccacaatc
     5221 tttcttgaaa gatgattcca tagacaataa agtattaact agaagtgata aaaatagagg
     5281 taaaagtgat aatgtcccaa gtgaggaagt cgtcaaaaag atgaaaaatt actggcgtca
     5341 acttttgaat gctaaattaa ttactcaaag aaaatttgat aatttgacta aagcagaaag
     5401 aggtgggctt tctgaattag ataaagccgg gttcattaaa agacaattgg tcgaaactag
     5461 acaaattact aaacatgttg cccaaatttt agattcccgt atgaacacta agtatgacga
     5521 aaatgataag ttaatacgtg aggttaaagt cattacttta aaatcaaaac ttgtctctga
     5581 tttcagaaag gatttccaat tctataaagt tagagaaatt aataattatc atcatgctca
     5641 tgatgcatat ttgaatgctg tagttggaac tgctttaatc aagaaatacc ctaaattaga
     5701 atctgaattt gtatatggtg attacaaagt ctatgatgtt agaaagatga ttgctaaatc
     5761 agaacaagaa attggtaaag ctacagctaa atacttcttt tactctaaca ttatgaattt
     5821 ctttaaaaca gaaattactt tggcaaacgg tgaaattaga aaaagacctc ttattgaaac
     5881 aaatggtgag actggagaga tagtttggga caaagggcgt gatttcgcta ctgttagaaa
     5941 agttttatca atgccacaag ttaacattgt aaagaaaaca gaggttcaaa ctggtggttt
     6001 ctcaaaagaa agtattttgc ctaaaagaaa tagtgataaa ttgattgcca gaaaaaagga
     6061 ttgggatcca aagaaatatg gtggtttcga ctcaccaacc gtagcctatt ctgttttggt
     6121 tgtggcaaag gttgaaaagg gtaaaagtaa aaagcttaaa tcagtaaaag aacttttggg
     6181 tattacaata atggaaagaa gttcctttga aaagaaccct attgattttt tggaagctaa
     6241 aggttataag gaagtaaaga aggacttaat aatcaaattg cctaaatatt ctttatttga
     6301 attagaaaat gggagaaaaa gaatgttggc ttctgctgga gaattgcaaa agggtaatga
     6361 attagcattg ccttccaaat atgttaactt cttgtattta gcttcacact atgaaaagtt
     6421 gaaagggtca ccagaagata acgagcaaaa acaattattt gttgaacaac acaaacacta
     6481 cttagatgag attatagaac aaattagtga attcagtaaa agagtgatat tagctgatgc
     6541 aaatttagat aaagttttgt cagcctataa caaacataga gataagccaa ttagagaaca
     6601 agcagaaaac attattcact tatttaccct taccaattta ggagcacctg ctgctttcaa
     6661 gtattttgat acaacaattg atcgtaaaag atatacctca acaaaagaag tcttagacgc
     6721 caccttaatt catcaatcaa tcactggatt gtatgagaca agaattgatt tgtctcaatt
     6781 gggtggtgat gaaggggctg atcctaagaa gaaaagaaaa gttgatccaa agaaaaagcg
     6841 taaggtggat cctaagaaaa agagaaaggt tgactacaaa gaccatgacg gtgattataa
     6901 agatcatgac atcgactaca aggatgacga tgacaagtga taatgactgc agagatccat
     6961 cgacctgccg ccaagctaat tccgggcgaa tttctgtcga gtcatgtaat tagttatgtc
     7021 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     7081 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     7141 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     7201 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccgggcccc
     7261 cctcgaggaa gttcctatac tttctagaga ataggaactt cggatccact agttctagat
     7321 ttttgcaagc atttaaatat tgccaagtaa aaacttcaaa ttttctttcc ccttggaact
     7381 ttgactttat ttttttgaca gattattttg acacacacac accaaatgtg ttacccctta
     7441 aaacaaaaaa acactttttt acaatttctt ggtatccaga atcattctaa gcatcattca
     7501 attataattt caatccaaaa aagtagtttt agtttgactt gaaacgtcaa caaacacaaa
     7561 tttcaaatca taacctctcc tgttgcctgt caacaacaca ccataaggag aaggaatagg
     7621 aggaggagga gatagaaact tgcacggcac cacaaaacac aaaattgatt tcaaccaata
     7681 cggtgacaac aacaatagat ttccgataga aataatgatt atcggaataa gctagctttg
     7741 ctttgctttg ctttgctttt tgacttgctc taatttttcg aaaataataa tggagaaaag
     7801 ttcaaggtgt ttaatgcatc aactaaaaca gaaaataata cattagacta aacttttaat
     7861 ctttctagta ccaataattc acgcgtgcgt tttaatccca atcatgaaat gaagaagtta
     7921 tttccctttt tctttcatca aaaaagaact aaattatttt ttaaatttta gtaaacaaaa
     7981 cctggaaatc ggggaaaccg ggggaggggg gcagaaggtg aaacgggtaa tattgataaa
     8041 tttaatctat aattgataaa gttaaattta aattgatttg aattgatttg aattgaatga
     8101 aatgcatttg aataaacggc atcaaactaa aaaaatatag atcacattca tagtaaaacg
     8161 ataacaaaga acaccacaat ttatagcaat gataataaac atctaaaaag aaaagggtac
     8221 gagaaggaga atgaaaaaaa acaataagct agttcttaat ctgttcagat atctaatttc
     8281 aaaaaaaaga atagtataaa aggatagttg attcctcttg gttgttgaaa atttgaataa
     8341 tatcaatcaa ttaatcaatc aaataacaac aacccactag acatcaccat tgtcgacatg
     8401 ccacaatttg atatattatg taaaacacca cctaaggtgc ttgttcgtca gtttgtggaa
     8461 aggtttgaaa gaccttcagg tgagaaaata gcattatgtg ctgctgaact aacctattta
     8521 tgttggatga ttacacataa cggaacagca atcaagagag ccacattcat gagctataat
     8581 actatcataa gcaattcgtt gagtttcgat attgtcaata aatcactcca gtttaaatac
     8641 aagacgcaaa aagcaacaat tttggaagcc tcattaaaga aattgattcc tgcttgggaa
     8701 tttacaatta ttccttacta tggacaaaaa catcaatctg atatcactga tattgtaagt
     8761 agtttgcaat tacagttcga atcatcggaa gaagcagata agggaaatag ccacagtaaa
     8821 aaaatgctta aagcacttct aagtgagggt gaaagcatct gggagatcac tgagaaaata
     8881 ctaaattcgt ttgagtatac ttcgagattt acaaaaacaa aaactttata ccaattcctc
     8941 ttcctagcta ctttcatcaa ttgtggaaga ttcagcgata ttaagaacgt tgatccgaaa
     9001 tcatttaaat tagtccaaaa taagtatttg ggagtaataa tccagtgttt agtgacagag
     9061 acaaagacaa gcgttagtag gcacatatac ttctttagcg caaggggtag gatcgatcca
     9121 cttgtatatt tggatgaatt tttgaggaat tctgaaccag tcctaaaacg agtaaatagg
     9181 accggcaatt cttcaagcaa taaacaggaa taccaattat taaaagataa cttagtcaga
     9241 tcgtacaata aagctttgaa gaaaaatgcg ccttattcaa tctttgctat aaaaaatggc
     9301 ccaaaatctc acattggaag acatttgatg acctcatttc tttcaatgaa gggcctaacg
     9361 gagttgacta atgttgtggg aaattggagc gataagcgtg cttctgccgt ggccaggaca
     9421 acgtatactc atcagataac agcaatacct gatcactact tcgcactagt ttctcggtac
     9481 tatgcatatg atccaatatc aaaggaaatg atagcattga aggatgagac taatccaatt
     9541 gaggagtggc agcatataga acagctaaag ggtagtgctg aaggaagcat acgatacccc
     9601 gcatggaatg ggataatatc acaggaggta ctagactacc tttcatccta cataaataga
     9661 cgcatataag agtgaaattc tggaaatctg gaaatctggt tttgtattct tgttattctt
     9721 ctttttgtta ttacatatat aacttgttac ttttttaaaa aaatctttgt atattttata
     9781 aatatataaa actaaattta agaaaaagag aaaaatgttt tatttgagag attgaaattt
     9841 tacttgaatt tagcttagct tttataaagt attattatgt aaaaaaacaa aacaaatata
     9901 cattaaaaag ttaagactat aaaatagcca cccaaggcat ttctatatct tgttgttgtt
     9961 gttttcatct tctgtatcag aggaacttat tttattattt tcgtcacggg tattttctct
    10021 tgtttgatga ttcatcccat tcattccatc ataaaatgtc gacactggat ggcggcgtta
    10081 gtatcgaatc gacagcagta tagcgaccag cattcacata cgattgacgc atgatattac
    10141 tttctgcgca cttaacttcg catctgggca gatgatgtcg aggcgaaaaa aaatataaat
    10201 cacgctaaca tttgattaaa atagaacaac tacaatataa aaaaactata caaatgacaa
    10261 gttcttgaaa acaagaatct ttttattgtc agtactgact cgagttatta tggacatggc
    10321 atagacatat acaaagcttg ttcaccatcg gaagcagtac catcgtataa agcagtatcc
    10381 aaaccacaca aagtgaaacc cattcttcta taagcatgaa tagctggagc attaacattg
    10441 gtaacttcca accacaaatg accagcacct ctttctctgg cgaattcagt agccaaaccc
    10501 atcaaagctc taccaacacc atgacctcta tgttctggag caacttcaat atcttcaaca
    10561 gtcaatcttc tgttccaacc agaataagaa acaacaacga aaccagccaa atcaccatca
    10621 tcaccataag caacgaaagt tctagaatct ggatcaccat cttcaccagc atcggattca
    10681 tcatcggatt catcatctgg gaaaacctta gtcaatggtg gatcaactgg aacttctctc
    10741 aaagtgaaac catcaccagt agcagtaact ctaaaaacag tatcggtagt gaaagaacca
    10801 tccaaagctt caatagcttc agcatcacct ggaacagaag ttctgtatct ataagcagta
    10861 tcatccaaag tagtagacat aattgtagga tccggttgtt tatgttcgga tgtgatgtga
    10921 gaactgtatc ctagcaagat tttaaaagga agtatatgaa agaagaacct cagtggcaaa
    10981 tcctaacctt ttatatttct ctacaggggc gcggcgtggg gacaattcaa cgcgtctgtg
    11041 aggggagcgt ttccctgctc gcaggtctgc agcgaggagc cgtaattttt gcttcgcgcc
    11101 gtgcggccat caaaatgtat ggatgcaaat gattatacat ggggatgtat gggctaaatg
    11161 tacgggcgac agtcacatca tgcccctgag ctgcgcacgt caagactgtc aaggagggta
    11221 ttctgggcct ccatgtcgct ggccgggtga cccggcgggg acgaggcaag cttgatggaa
    11281 gttcctatac tttctagaga ataggaactt cagatccact agttctagag cggccgccac
    11341 cgcggcgaag aagaattagg ttctgaagct atctacgctg gtaaagattt ccaaaaggct
    11401 tctcaattgt aagtttgcct ctgattaaat aaaaaaaagc tggtgctttt ttttctttta
    11461 taggaacatc ttgaatatat gaactaatta aatgataatt ttttacccat ctttactctt
    11521 aatcactgag ctgcagtcaa agaaaaaggg atacagcacc tggtgaagag atgaacggag
    11581 actaacttag acgcgttgat tcttttcaat tgcacatttt attaatcgat gctaacgtct
    11641 atttacatat attctttaga gatattatct agggcttcaa ataatctctg gacagcaata
    11701 aaagtctctt caaaagtatt gtataacggc aatggggcta atctgattac atctggtctt
    11761 ctttcgtcac agattatagc atgatcatgc aagtacgcat taactcgttc catgacgttc
    11821 ttgccctttt catcgaaatg cggttgaaac ataatggaca attgacatcc tctttcagct
    11881 ggattcaaag gagttaaaat tttaaaccca aatttggagt ttgatgtact ggattgtggt
    11941 atgtaatact tggaattcgt caatagatcc tgtaaaaatt gagtcaaagc aacacttttt
    12001 tcacgaagtt tagatactcc acccacttta gcatacactt ccaatgacga cttcacagca
    12061 acaacatcaa gaacagaagg atttgactgt ctgtaagaaa gagccgagtt tattggatca
    12121 aactcttcta acattttgaa tcgttcttgg gagttattgc cccaccaacc agctagtcta
    12181 ggaacgaaac tgctcttctt gttctctata gtgtattttt catgcacaaa aattccacct
    12241 atggctccag gtcccgagtt taaatatttg taggaacacc aagcagcaaa atctactccc
    12301 caatcatgta aatttaatgg aacattccca actgcatggg caagatccca cccaacttta
    12361 atttgttggc tcttttcctt agcgtattta gttatttcct ctatcttgaa aaattgacca
    12421 gtgtagtatt ggataccagg aaaacacact agagccaatt catccaggtt ctcatctata
    12481 gccttgatta ttctttctgt tttaatataa gtttcaccag gttgaacttc caattgaatc
    12541 aaatgtttct cgtcgtatcc gaacaattta acaatgttca aaaatgcata gtagtcagaa
    12601 ggaaatgctt gtttttcaaa taaaattttg gttcttttcc cctcaggttt gtaaaaatgg
    12661 atcaacaatg cattcaagtt tgctgttaaa gaacccataa ctgcaacttc gttttccttt
    12721 gcaccaacaa tgggggctat taatggtaat aagggtaaat cgatgtctac ccacggtgtt
    12781 aacagtttgt caggatgatt gaaatgagac tcaacccctc gttcaaccca tgcattcaat
    12841 tcatcattga tagctttctt tgtattctta ggcatcaacc caagagagtt tccacataaa
    12901 taaatagact cagttgatga ctcatattta ttatttttga tacctaatga tccaaaagtt
    12961 ggtatggcaa actcattttt aaaagttggg aactttttgt ccaatttctt tgcctcggct
    13021 aatgacatct gataataaaa tggggttgga gtagttggtg gtataaccgg agagatagaa
    13081 ttgaagaaaa aaatcggaaa caacaaaaaa agttgatacc cagtattatg tgggagataa
    13141 ttgcgaatgg tggaaaaaaa aaagacgcct ttgggcctca acaacaattc taacagcttg
    13201 agagctttac aatcgagaaa ctatgattca ttccgtttta atatgtatgt acttagtaaa
    13261 ctcgtgaatt ttatttgtgg tctactttag tactaacata atcattggat agtcaataat
    13321 gatggtcttc cgagactaat gaaattctat accaaagtcg atattccaac acagaaattg
    13381 ctcttgcaac aagtgcacct gttgatatct actagatcag cacaagagtg ttcggagctc
    13441 gcatgcaggc ctcagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
    13501 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
    13561 agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
    13621 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
    13681 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
    13741 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
    13801 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
    13861 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
    13921 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
    13981 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
    14041 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
    14101 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
    14161 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
    14221 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
    14281 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
    14341 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
    14401 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
    14461 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
    14521 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
    14581 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
    14641 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
    14701 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
    14761 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
    14821 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
    14881 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
    14941 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
    15001 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
    15061 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
    15121 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
    15181 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
    15241 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    15301 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    15361 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    15421 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    15481 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    15541 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    15601 ttagaaaaat aaacaaatag gggttccgcg cacattt
//
