LOCUS       Exported               14105 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  CaCas9/gRNA Solo entry plasmid for cloning guides - contains stuffer
            with BsmBI sites - integrates at ENO1.
ACCESSION   .
VERSION     .
KEYWORDS    pV1093
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14105)
  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 14105)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14105
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(15..470)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(152..173)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     364..383
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     597..619
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     611..628
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     612..628
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2243..6346
                     /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             6359..6379
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6383..6403
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6407..6427
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6428..6493
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      6558..6805
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6574..6592)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     misc_RNA        9722..9797
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(11929..11949)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11929..11947)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11968..11984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11968..11984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11981..12003)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11992..12008
                     /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(12016..12046)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12061..12082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12199..12216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12370..12958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12450..12469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13129..13989)
                     /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     13752..13771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13990..14094)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
       61 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
      121 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
      181 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
      241 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
      301 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
      361 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
      421 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
      481 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
      541 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
      601 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
      661 attgaggcct gcatgcggta cctgccacta ctaccactgg gagtttcgtt cttctcgata
      721 ctattagctt tacttcctgc actagcagtg gttggatcaa cagaatcttc ataatcatca
      781 aaatcgtctt ttgaagaccc cccgtttgat gtatggccct gtcttttcat caaacttttt
      841 atatagttga ctgaactgag gctaaatatg tgatcatctt cactatagac aatctttctc
      901 ttatttgcac caccgccacc accagtcttt gagaaattct caaaaccttt tacgatatta
      961 ccaagcgggc tctcttcgaa ataatctatc tctttttgat atatcgaatc ctctagcgtg
     1021 gttagctttc tagttagttc ttgcttctta agaatttgct ggattagttt atttttcaat
     1081 tcaacgtatt tctcagagtc atctttagat tttgatgaag atgtgcgttc attcgctata
     1141 tccttcttgg tcgtgtcttt tcgatcctcc ttggctggca ctgaactcgt cttttttggc
     1201 gttgctgttc cagacagact tatctcatta gatttggaac ttgtgggttt aacatcattt
     1261 gcatctttag tagacatgat tgtgtaatac cgtgattgtt tgttttgaaa ggtctgtcat
     1321 atttctatca atttcaaaac aaaatgttca tcagaaaaaa gccaaaaatg tctcttctag
     1381 cttcttagtg gtgtcgcaaa atacacaatg tcgctcaaca atccacattc ccggcgcata
     1441 gctcaaatca catgactaca gctaacaatt acacaaaaaa attctctttt cgatgtagca
     1501 actatcttca actaaaacat tttctccttc ggcccatgat tgtcctccgg gtcgacagca
     1561 agccgttaca attgagatgg aaagcgacct accttcactc gataaggtgc ttaattgtac
     1621 ttcatataaa tctggcccgg atctaaacaa atgagttcca ttaagccgtg ggttctcaat
     1681 tagggttttt gtttttgatt tagaaaaaag agatcaagat ttgtttacag gtgatgcctt
     1741 tttttagaac ttatgcgttg caaaagttga ctaacgattt ctataaggtg atccacacta
     1801 attatacaaa cgtacaaaca gacatacttt tcctgcgttc acctgatgtt ggccagattt
     1861 ctctcttcat tgcatagaac ataaccacac tagggcaaca gnaaaaaaaa aaaaaaagtg
     1921 catcgggaag ttgtgttcca ttcattatat gtctactact gcatatgagt agcccaccca
     1981 ccaccaccat agtaagtttt tgtgtatgcg cgccgtcagg ttatttcatt tctgaatttt
     2041 tcaaccacct tactcccttt attgttgatt gacaattttg ctcacagtaa gatcttttag
     2101 actccaatta atataaaata agtctgattt tccaattcct gttttttctt tttttttctg
     2161 tttctatttc tttccttttc tccctttttt ttaattcttc attcaatcat caattgataa
     2221 ttcaggaata ttacaacccg ggatggataa aaagtatagt attggtttag atattggtac
     2281 taactctgtg ggttgggcag ttatcactga cgaatataaa gttccatcaa agaaatttaa
     2341 ggtgttaggt aacactgaca gacactcaat aaaaaagaat cttatcggtg ctcttttgtt
     2401 cgactccggt gaaactgccg aggctacacg tttaaaaaga acagcaagaa gaagatatac
     2461 ccgtagaaaa aatagaatat gttatttaca agaaatcttt tctaatgaaa tggctaaagt
     2521 tgatgattcc tttttccata gattggaaga gtcatttttg gttgaagaag acaaaaagca
     2581 tgagagacat ccaatctttg ggaatatagt tgatgaagtg gcttaccatg aaaaatatcc
     2641 taccatttat catttaagaa agaaattggt agattcaact gataaagctg accttagatt
     2701 aatctattta gcacttgccc atatgattaa atttagaggt cattttttga ttgaaggtga
     2761 tttgaaccca gataattctg acgtggataa attatttatt caattagtcc aaacctacaa
     2821 ccaattattt gaggaaaatc caattaatgc tagtggtgtc gatgccaaag ctatattatc
     2881 agccagatta tcaaaatcta gacgtttgga aaatttgatt gcccaattgc caggagaaaa
     2941 aaagaatgga ttatttggaa acttgatcgc attatcattg ggtttgacac caaattttaa
     3001 atctaatttt gatttagctg aagatgctaa attacaatta tcaaaagaca cctatgacga
     3061 cgatttggac aatttacttg ctcaaattgg tgatcaatat gcagatttgt tcttagctgc
     3121 taaaaactta tctgatgcta ttttgttgtc tgatattttg agagtgaaca cagaaataac
     3181 caaagctcca ttatcagcat ctatgatcaa acgttatgat gaacaccatc aggatttgac
     3241 tttattgaaa gctttggtga gacaacaatt gccagagaag tataaagaaa tctttttcga
     3301 tcaatctaaa aacgggtatg caggttatat tgatgggggt gcctcccaag aggaatttta
     3361 caaatttata aaacctattt tagaaaagat ggatgggact gaggaacttt tggtcaaatt
     3421 gaacagagaa gatttgttac gtaaacagag aacttttgat aatggtagta tacctcacca
     3481 aattcatttg ggtgagttgc atgcaatttt aagaagacaa gaagattttt atccattttt
     3541 aaaagataat agagaaaaaa tcgagaaaat tttaaccttt agaattccat actatgttgg
     3601 gcctttggct agaggtaatt caagatttgc ctggatgaca cgtaaatcag aagaaactat
     3661 taccccttgg aattttgaag aggttgttga taaaggagca tcagcacaga gttttattga
     3721 aagaatgacc aatttcgata aaaacttacc aaatgaaaaa gttttaccaa aacattcctt
     3781 gttatacgaa tattttactg tttacaatga acttacaaag gttaaatatg ttactgaagg
     3841 tatgcgtaag ccagcctttt tatctggaga acagaaaaag gcaatagttg atttattgtt
     3901 taaaacaaat agaaaagtta ctgttaaaca attaaaagaa gattacttta agaaaattga
     3961 atgttttgat tcagttgaaa tcagtggtgt tgaagacaga tttaatgcta gtttaggaac
     4021 ttaccatgat ttacttaaaa ttatcaaaga taaagatttc ttggataacg aagaaaatga
     4081 agacatttta gaagacattg ttttaacctt aactttattc gaagatagag agatgattga
     4141 agaacgtttg aagacttatg cacatttgtt tgacgataaa gtgatgaaac agttgaaaag
     4201 aagacgttat actggatggg gtagattgtc tcgtaaattg atcaatggaa ttagagataa
     4261 acaaagtggt aaaactatct tggacttttt gaaatctgac ggatttgcta atagaaattt
     4321 catgcaattg atccacgacg atagtttgac atttaaagaa gacatccaaa aggcccaagt
     4381 gagtgggcaa ggtgattcat tacatgaaca tattgcaaat ttagccggat ctcctgctat
     4441 taagaaaggg atattacaaa ctgttaaagt tgtggatgaa ttagtgaaag taatgggaag
     4501 acataaacct gaaaacattg tcattgagat ggcaagagaa aatcaaacta cacaaaaagg
     4561 acagaaaaat agtagagaac gtatgaaaag aatagaagag ggtattaaag aattgggtag
     4621 tcaaatattg aaagaacacc cagtggaaaa tacccagttg caaaatgaaa aattatatct
     4681 ttactacctt caaaatggac gtgatatgta tgttgatcag gaattagata taaatagact
     4741 ttcagattat gatgtagatc atatagttcc acaatctttc ttgaaagatg attccataga
     4801 caataaagta ttaactagaa gtgataaaaa tagaggtaaa agtgataatg tcccaagtga
     4861 ggaagtcgtc aaaaagatga aaaattactg gcgtcaactt ttgaatgcta aattaattac
     4921 tcaaagaaaa tttgataatt tgactaaagc agaaagaggt gggctttctg aattagataa
     4981 agccgggttc attaaaagac aattggtcga aactagacaa attactaaac atgttgccca
     5041 aattttagat tcccgtatga acactaagta tgacgaaaat gataagttaa tacgtgaggt
     5101 taaagtcatt actttaaaat caaaacttgt ctctgatttc agaaaggatt tccaattcta
     5161 taaagttaga gaaattaata attatcatca tgctcatgat gcatatttga atgctgtagt
     5221 tggaactgct ttaatcaaga aataccctaa attagaatct gaatttgtat atggtgatta
     5281 caaagtctat gatgttagaa agatgattgc taaatcagaa caagaaattg gtaaagctac
     5341 agctaaatac ttcttttact ctaacattat gaatttcttt aaaacagaaa ttactttggc
     5401 aaacggtgaa attagaaaaa gacctcttat tgaaacaaat ggtgagactg gagagatagt
     5461 ttgggacaaa gggcgtgatt tcgctactgt tagaaaagtt ttatcaatgc cacaagttaa
     5521 cattgtaaag aaaacagagg ttcaaactgg tggtttctca aaagaaagta ttttgcctaa
     5581 aagaaatagt gataaattga ttgccagaaa aaaggattgg gatccaaaga aatatggtgg
     5641 tttcgactca ccaaccgtag cctattctgt tttggttgtg gcaaaggttg aaaagggtaa
     5701 aagtaaaaag cttaaatcag taaaagaact tttgggtatt acaataatgg aaagaagttc
     5761 ctttgaaaag aaccctattg attttttgga agctaaaggt tataaggaag taaagaagga
     5821 cttaataatc aaattgccta aatattcttt atttgaatta gaaaatggga gaaaaagaat
     5881 gttggcttct gctggagaat tgcaaaaggg taatgaatta gcattgcctt ccaaatatgt
     5941 taacttcttg tatttagctt cacactatga aaagttgaaa gggtcaccag aagataacga
     6001 gcaaaaacaa ttatttgttg aacaacacaa acactactta gatgagatta tagaacaaat
     6061 tagtgaattc agtaaaagag tgatattagc tgatgcaaat ttagataaag ttttgtcagc
     6121 ctataacaaa catagagata agccaattag agaacaagca gaaaacatta ttcacttatt
     6181 tacccttacc aatttaggag cacctgctgc tttcaagtat tttgatacaa caattgatcg
     6241 taaaagatat acctcaacaa aagaagtctt agacgccacc ttaattcatc aatcaatcac
     6301 tggattgtat gagacaagaa ttgatttgtc tcaattgggt ggtgatgaag gggctgatcc
     6361 taagaagaaa agaaaagttg atccaaagaa aaagcgtaag gtggatccta agaaaaagag
     6421 aaaggttgac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     6481 tgacgatgac aagtgataat gactgcagag atccatcgac ctgccgccaa gctaattccg
     6541 ggcgaatttc tgtcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc
     6601 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     6661 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     6721 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     6781 gggacgctcg aaggctttaa tttgcggccg ggccccccct aactcaagta caacagatct
     6841 ggaccacctt tgattgtaaa tagtaataat taccaccctt atctaattat ttatttaact
     6901 tatttattta tttattatac atatatacaa acctaataaa gtgaaaatct cccccttcac
     6961 acttcacata tgttaggcgt catcctgtgc tcccgagaac cagtaccagt acatcgctgt
     7021 ttcgttcgag acttgaggtc tagttttata cgtgaagagg tcaatgccgc cgagagtaaa
     7081 gccacatttt gcgtacaaat tgcaggcagg tacattgttc gtttgtgtct ctaatcgtat
     7141 gccaaggagc tgtctgctta gtgcccactt tttcgcaaat tcgatgagac tgtgcgcgac
     7201 tcctttgcct cggtgcgtgt gcgacacaac aatgtgttcg atagaggcta gatcgttcca
     7261 tgttgagttg agttcaatct tcccgacaag ctcttggtcg atgaatgcgc catagcaagc
     7321 agagtcttca tcagagtcat catctgagat gtaatccttc cggtaggggc tcacacttct
     7381 ggtagatagt tcaaagcctt ggtcggatag gtgcacatcg aacacttcac gaacaatgaa
     7441 atggttctca gcatccaatg tttccgccac ctgctcaggg atcaccgaaa ttttcatatg
     7501 agaaccgtta tcgataacta aagcagcaac ttcttctata aaaatgggtt agtatgacag
     7561 tcatttaaat aaggaatttt tcagttggct tggtttcaat tcaatgttcg tttttttttt
     7621 tcttgctgtg tttgtgtttg tgttgtttat agttgtgtgc actgatcgtc gaaaaaaaaa
     7681 attcatagtg agccgggaaa tctgtatagc ccagataaca acacaagtcc aaactagaaa
     7741 ctcgtcaaac accaaaagca atgttgaatc aattgccttg cacaagtaca cgtaggaaaa
     7801 cataaaacat tgcaattttg aatattgagc cttttgtcgt aacattgatt gataggatta
     7861 ctcaccgaat ggttttgaaa ccactgccga cagatcaatc aatcaatcaa aaaacgtgaa
     7921 ctttgaaaaa ggggaagaac agatacattg aagttagcca tttccactga tcgtcacaac
     7981 atatctgata aattactttc aaaattataa gctgatgtgt gtgtattatt aatgtgacag
     8041 taacatccca aacgagaaat attatctcga caacaaaaaa gtttgatctg aattgaaaat
     8101 gaagttttcc caccctaccc atttgtcata ttgaaaccaa tcaactgatt aatcaatcaa
     8161 ttagaattga agctaaacta aaacatacca ccgtccattt tgaatgatta tattttttta
     8221 atattaatat cgagataatg tttctaagaa agaaagaaaa ccaggagtga aaattagaaa
     8281 aggaaaggaa agaaaaaaag aaaaatctga aaatatataa aaaaaaattg tttcgttggc
     8341 aataaatctt ggtgagaaca gcgaccgaaa gcaaataaga acaaaatatg agtgtattac
     8401 gttgaacaac taattaacgt gtgtgtatgg atcttttttt cttttttctc tttaaccgac
     8461 tataaacaac aaacattttt gggcagtgca cacactactt aatatacaca gcataaatta
     8521 cacgattaga aacaaattag cttattaaaa taacctaatc aaaccgaata ttttatggta
     8581 ttatgagtaa actatataat ataaatagca cacacccaca acaacaacaa aggaaaacta
     8641 aaaggttttt tctttttgaa aagatcgttt tctttattat tctctagttt tgacggcggc
     8701 cgcagtgatt agacttagtc cgttcaaatc aagcacaact ctgttcattg tttcaacaag
     8761 aattaattca aaaacaggtt cggtgcataa tttgcaaaaa aatattgcag cttctgtggc
     8821 tcgaacacag tacctccaga tttcaggttt gaaatacttc agtctgacgc tctcccagat
     8881 gagctaaagc tgcaataaga aaacccacgc cgggattcga acccggaatc ctttgattag
     8941 aagtcaaaag cgataaccat ttcgccacgc aggcctactt gatgggtttg taaatggtct
     9001 actttttcag acctaacaga aattttaatg aaagtcatat tcttatacaa taaaactgtg
     9061 tcataaaagc agatattcga ctttcgtaga ttatatagga cccaagaact aaaatttaat
     9121 gccatattat gcatttttaa tctgtaaaag tgttgtttcc aacctatcac aagtacgttc
     9181 ttgtaacttg tgtttgtagg gttgcaaatg aatcataaca acatctcaac agaacatgta
     9241 tagcaaagct tagtataaaa tcagtgtttt gagaggcaat ccaagaatgt ttacatcaaa
     9301 gtttcaataa atatcgaccg aaactgaaaa tctttttagg ttattgttca cttttttgta
     9361 aatatttaaa cattttttgg acctaaaaaa atacaagcac caattacgta ccaagaagca
     9421 tctaatcaac tcccagatca ccactataca tttaaaagtc attggtcaat aactatactc
     9481 gagtattgcc tcatcaaaga aacaatcaaa tattatagat actcactcca tcacgtgata
     9541 atttcactgg tatggaaaag tggaaaattt tataaaaaaa aatttgatgc ctttggcata
     9601 gctgaaactt cggcccaata ggattggaga atatgttttc gcagcgttct tacaattaaa
     9661 ttgtggtgga agttcgagac ttgcgtaaac tatttttaat ttggagacgg aattccgtct
     9721 cgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     9781 tggcaccgag tcggtgcttt ttttctcgag tttttttatc gagtgtttaa ggataatgat
     9841 aactgaagag aagaattagt tttgccgcca ccgcgggttt gcctctgatt aaataaaaaa
     9901 aagctggtgc ttttttttct tttataggaa catcttgaat atatgaacta attaaatgat
     9961 aattttttac ccatctttac tcttaatcac tgagctgcag tcaaagaaaa agggatacag
    10021 cacctggtga agagatgaac ggagactaac ttagacgcgt tgattctttt caattgcaca
    10081 ttttattaat cgatgctaac gtctatttac atatattctt tagagatatt atctagggct
    10141 tcaaataatc tctggacagc aataaaagtc tcttcaaaag tattgtataa cggcaatggg
    10201 gctaatctga ttacatctgg tcttctttcg tcacagatta tagcatgatc atgcaagtac
    10261 gcattaactc gttccatgac gttcttgccc ttttcatcga aatgcggttg aaacataatg
    10321 gacaattgac atcctctttc agctggattc aaaggagtta aaattttaaa cccaaatttg
    10381 gagtttgatg tactggattg tggtatgtaa tacttggaat tcgtcaatag atcctgtaaa
    10441 aattgagtca aagcaacact tttttcacga agtttagata ctccacccac tttagcatac
    10501 acttccaatg acgacttcac agcaacaaca tcaagaacag aaggatttga ctgtctgtaa
    10561 gaaagagccg agtttattgg atcaaactct tctaacattt tgaatcgttc ttgggagtta
    10621 ttgccccacc aaccagctag tctaggaacg aaactgctct tcttgttctc tatagtgtat
    10681 ttttcatgca caaaaattcc acctatggct ccaggtcccg agtttaaata tttgtaggaa
    10741 caccaagcag caaaatctac tccccaatca tgtaaattta atggaacatt cccaactgca
    10801 tgggcaagat cccacccaac tttaatttgt tggctctttt ccttagcgta tttagttatt
    10861 tcctctatct tgaaaaattg accagtgtag tattggatac caggaaaaca cactagagcc
    10921 aattcatcca ggttctcatc tatagccttg attattcttt ctgttttaat ataagtttca
    10981 ccaggttgaa cttccaattg aatcaaatgt ttctcgtcgt atccgaacaa tttaacaatg
    11041 ttcaaaaatg catagtagtc agaaggaaat gcttgttttt caaataaaat tttggttctt
    11101 ttcccctcag gtttgtaaaa atggatcaac aatgcattca agtttgctgt taaagaaccc
    11161 ataactgcaa cttcgttttc ctttgcacca acaatggggg ctattaatgg taataagggt
    11221 aaatcgatgt ctacccacgg tgttaacagt ttgtcaggat gattgaaatg agactcaacc
    11281 cctcgttcaa cccatgcatt caattcatca ttgatagctt tctttgtatt cttaggcatc
    11341 aacccaagag agtttccaca taaataaata gactcagttg atgactcata tttattattt
    11401 ttgataccta atgatccaaa agttggtatg gcaaactcat ttttaaaagt tgggaacttt
    11461 ttgtccaatt tctttgcctc ggctaatgac atctgataat aaaatggggt tggagtagtt
    11521 ggtggtataa ccggagagat agaattgaag aaaaaaatcg gaaacaacaa aaaaagttga
    11581 tacccagtat tatgtgggag ataattgcga atggtggaaa aaaaaaagac gcctttgggc
    11641 ctcaacaaca attctaacag cttgagagct ttacaatcga gaaactatga ttcattccgt
    11701 tttaatatgt atgtacttag taaactcgtg aattttattt gtggtctact ttagtactaa
    11761 cataatcatt ggatagtcaa taatgatggt cttccgagac taatgaaatt ctataccaaa
    11821 gtcgatattc caacacagaa attgctcttg caacaagtgc acctgttgat atctactaga
    11881 tcagcacaag agtgttcgga gctcgcatgc aggcctcagc ttttgttccc tttagtgagg
    11941 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
    12001 gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta
    12061 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
    12121 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
    12181 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
    12241 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
    12301 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
    12361 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
    12421 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
    12481 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
    12541 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    12601 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    12661 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    12721 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    12781 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    12841 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    12901 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    12961 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
    13021 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
    13081 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
    13141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
    13201 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
    13261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
    13321 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
    13381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
    13441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
    13501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
    13561 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
    13621 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
    13681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
    13741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
    13801 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
    13861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
    13921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    13981 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
    14041 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
    14101 tcccc
//
