pFC332

LOCUS       pFC332               15559 bp ds-DNA     circular SYN 10-JAN-2024
DEFINITION  AMA1 plasmid with Aspergillus optimized Cas9 and hph selection 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    pFC332
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15559)
  AUTHORS   Nodvig CS, Nielsen JB, Kogle ME, Mortensen UH
  TITLE     A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi.
  JOURNAL   PLoS One. 2015 Jul 15;10(7):e0133085. doi: 
            10.1371/journal.pone.0133085. eCollection 2015.
  PUBMED    26177455
REFERENCE   2  (bases 1 to 15559)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 10, 2024 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..15559
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1244..1277
                     /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)"
     rep_origin      1590..6837
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     CDS             8009..12112
                     /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             12116..12136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12962..13987
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(14059..14092)
                     /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(14562..14579)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(14562..14578)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14677..14694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14848..15436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14928..14947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(15558..859)
                     /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"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcgctacat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttat tcgactctat agtgaagttc ctattctcta
     1261 gaaagtatag gaacttctga agtggggatt taaatgcggc cgcgctgagg gtttaatcga
     1321 cgaagcagct gacggccagt gccaagctta acgcgtaccg ggcccagtat atgttccgca
     1381 gatgactgga gctctgccat acgtgccctc tcaagcacca tttgttccat ctacagagac
     1441 tagtcaccaa ctagtctatc aagactcaca gggtacattg ctgagaccaa ctgaccagag
     1501 gcagggtagc ggattgacgg ctccatctcc ttcacttaca aggtctattg aaagcccttt
     1561 agcatcacca agcggagaat agattgttaa gcttattttt tgtatactgt tttgtgatag
     1621 cacgaagttt ttccacggta tcttgttaaa aatatatatt tgtggcgggc ttacctacat
     1681 caaattaata agagactaat tataaactaa acacacaagc aagctacttt agggtaaaag
     1741 tttataaatg cttttgacgt ataaacgttg cttgtattta ttattacaat taaaggtgga
     1801 tagaaaacct agagactagt tagaaactaa tctcaggttt gcgttaaact aaatcagagc
     1861 ccgagaggtt aacagaacct agaaggggac tagatatccg ggtagggaaa caaaaaaaaa
     1921 aacaagacag ccacatatta gggagactag ttagaagcta gttccaggac taggaaaata
     1981 aaagacaatg ataccacagt ctagttgaca actagataga ttctagattg aggccaaagt
     2041 ctctgagatc caggttagtt gcaactaata ctagttagta tctagtctcc tataactctg
     2101 aagctagaat aacttactac tattatcctc accactgttc agctgcgcaa acggagtgat
     2161 tgcaaggtgt tcagagacta gttattgact agtcagtgac tagcaataac taacaaggta
     2221 ttaacctacc atgtctgcca tcaccctgca cttcctcggg ctcagcagcc ttttcctcct
     2281 cattttcatg ctcattttcc ttgtttaaga ctgtgactag tcaaagacta gtccagaacc
     2341 acaaaggaga aatgtcttac cactttcttc attgcttgtc tcttttgcat tatccatgtc
     2401 tgcaactagt tagagtctag ttagtgacta gtccgacgag gacttgcttg tctccggatt
     2461 gttggaggaa ctctccaggg cctcaagatc cacaacagag ccttctagaa gactggtcaa
     2521 taactagttg gtctttgtct gagtctgact tacgaggttg catactcgct ccctttgcct
     2581 cgtcaatcga tgagaaaaag cgccaaaact cgcaatatgg ctttgaacca cacggtgctg
     2641 agactagtta gaatctagtc ccaaactagc ttggatagct tacctttgcc ctttgcgttg
     2701 cgacaggtct tgcagggtat ggttcctttc tcaccagctg atttagctgc cttgctaccc
     2761 tcacggcgga tctgcataaa gagtggctag aggttataaa ttagcactga tcctaggtac
     2821 ggggctgaat gtaacttgcc yttcctttct catcgcgcgg caagacaggc ttgctcaaat
     2881 tcctaccagt cacaggggta tgcacggcgt acggaccact tgaactagtc acagattagt
     2941 tagcaactag tctgcattga atggctgtac ttacgggccc tcgccattgt cctgatcatt
     3001 tccagcttca ccctcgttgc tgcaaagtag ttagtgacta gtcaaggact agttgaaatg
     3061 ggagaagaaa ctcacgaatt ctcgacaccc ttagtattgt ggtccttgga cttggtgctg
     3121 ctatatatta gctaatacac tagttagact cacagaaact tacgcagctc gcttgcgctt
     3181 cttggtagga gtcggggttg ggagaacagt gccttcaaac aagccttcat accatgctac
     3241 ttgactagtc agggactagt caccaagtaa tctagatagg acttgccttt ggcctccatc
     3301 agttccttca tagtgggagg tccattgtgc aatgtaaact ccatgccgtg ggagttctta
     3361 tccttcaagt gcttgaccaa tatgtttctg ttggcagagg gaacctgtca actagttaat
     3421 aactagtcag aaactagtat agcagtagac tcactgtacg cttgaggcat cccttcactc
     3481 ggcagtagac ttcatatgga tggatatcag gcacgccatt gtcgtcctgt ggactagtca
     3541 gtaactaggc ttaaagctag tcgggtcggc ttactatctt gaaatccggc agcgtaagct
     3601 ccccgtcctt aactgcctcg agatagtgac agtactctgg ggactttcgg agatcgttat
     3661 cgcgaatgct cggcatacta atcgttgact agtcttggac tagtcccgag caaaaaggat
     3721 tggaggagga ggaggaaggt gagagtgaga caaagagcga aataagagct tcaaaggcta
     3781 tctctaagca gtatgaaggt taagtatcta gttcttgact agatttaaaa gagatttcga
     3841 ctagttatgt acctggagtt tggatatagg aatgtgttgt ggtaacgaaa tgtaaggggg
     3901 aggaaagaaa aagtcggtca agaggtaact ctaagtcggc cattcctttt tgggaggcgc
     3961 taaccataaa cggcatggtc gacttagagt tagctcaggg aatttaggga gttatctgcg
     4021 accaccgagg aacggcggaa tgccaaagaa tcccgatgga gctctagctg gcggttgaca
     4081 accccacctt ttggcgtttc tgcggcgttg caggcgggac tggatacttc gtagaaccag
     4141 aaaggcaagg cagaacgcgc tcagcaagag tgttggaagt gatagcatga tgtgccttgt
     4201 taactaggtc aaaatctgca gtatgcttga tgttatccaa agtgtgagag aggaaggtcc
     4261 aaacatacac gattgggaga gggcctaggt ataagagttt ttgagtagaa cgcatgtgag
     4321 cccagccatc tcgaggagat taaacacggg ccggcatttg atggctatgt tagtacccca
     4381 atggaaacgg tgagagtcca gtggtcgcag ataactccct aaattccctg agctaactct
     4441 aagtcgacca tgccgtttat ggttagcgcc tcccaaaaag gaatggccga cttagagtta
     4501 cctcttgacc gactttttct ttcctccccc ttacatttcg ttaccacaac acattcctat
     4561 atccaaactc caggtacata actagtcgaa atctctttta aatctagtca agaactagat
     4621 acttaacctt catactgctt agagatagcc tttgaagctc ttatttcgct ctttgtctca
     4681 ctctcacctt cctcctcctc ctccaatcct ttttgctcgg gactagtcca agactagtca
     4741 acgattagta tgccgagcat tcgcgataac gatctccgaa agtccccaga gtactgtcac
     4801 tatctcgagg cagttaagga cggggagctt acgctgccgg atttcaagat agtaagccga
     4861 cccgactagc tttaagccta gttactgact agtccacagg acgacaatgg cgtgcctgat
     4921 atccatccat atgaagtcta ctgccgagtg aagggatgcc tcaagcgtac agtgagtcta
     4981 ctgctatact agtttctgac tagttattaa ctagttgaca ggttccctct gccaacagaa
     5041 acatattggt caagcacttg aaggacaaga actcccacgg catggagttt acattgcaca
     5101 atggacctcc cactatgaag gaactgatgg aggccaaagg caagtcctat ctagattact
     5161 tggtgactag tccctgacta gtcaagtagc atggtatgaa ggcttgtttg aaggcactgt
     5221 tctcccaacc ccgactccta ccaagaagcg caagcgagct gcgtaagttt ctgtgagtct
     5281 aactagtgta ttagctaata tatagcagca ccaagtccaa ggaccacaat actaagggtg
     5341 tcgagaattc gtgagtttct tctcccattt caactagtcc ttgactagtc actaactact
     5401 ttgcagcaac gagggtgaag ctggaaatga tcaggacaat ggcgagggcc cgtaagtaca
     5461 gccattcaat gcagactagt tgctaactaa tctgtgacta gttcaagtgg tccgtacgcc
     5521 gtgcataccc ctgtgactgg taggaatttg agcaagcctg tcttgccgcg cgatgagaaa
     5581 ggaarggcaa gttacattca gccccgtacc taggatcagt gctaatttat aacctctagc
     5641 cactctttat gcagatccgc cgtgagggta gcaaggcagc taaatcagct ggtgagaaag
     5701 gaaccatacc ctgcaagacc tgtcgcaacg caaagggcaa aggtaagcta tccaagctag
     5761 tttgggacta gattctaact agtctcagca ccgtgtggtt caaagccata ttgcgagttt
     5821 tggcgctttt tctcatcgat tgacgaggca aagggagcga gtatgcaacc tcgtaagtca
     5881 gactcagaca aagaccaact agttattgac cagtcttcta gaaggctctg ttgtggatct
     5941 tgaggccctg gagagttcct ccaacaatcc ggagacaagc aagtcctcgt cggactagtc
     6001 actaactaga ctctaactag ttgcagacat ggataatgca aaagagacaa gcaatgaaga
     6061 aagtggtaag acatttctcc tttgtggttc tggactagtc tttgactagt cacagtctta
     6121 aacaaggaaa atgagcatga aaatgaggag gaaaaggctg ctgagcccga ggaagtgcag
     6181 ggtgatggca gacatggtag gttaatacct tgttagttat tgctagtcac tgactagtca
     6241 ataactagtc tctgaacacc ttgcaatcac tccgtttgcg cagctgaaca gtggtgagga
     6301 taatagtagt aagttattct agcttcagag ttataggaga ctagatacta actagtatta
     6361 gttgcaacta acctggatct cagagacttt ggcctcaatc tagaatctat ctagttgtca
     6421 actagactgt ggtatcattg tcttttattt tcctagtcct ggaactagct tctaactagt
     6481 ctccctaata tgtggctgtc ttgttttttt tttttgtttc cctacccgga tatctagtcc
     6541 ccttctaggt tctgttaacc tctcgggctc tgatttagtt taacgcaaac ctgagattag
     6601 tttctaacta gtctctaggt tttctatcca cctttaattg taataataaa tacaagcaac
     6661 gtttatacgt caaaagcatt tataaacttt taccctaaag tagcttgctt gtgtgtttag
     6721 tttataatta gtctcttatt aatttgatgt aggtaagccc gccacaaata tatattttta
     6781 acaagatacc gtggaaaaac ttcgtgctat cacaaaacag tatacaaaaa ataagcttaa
     6841 caatctattc tccgcttggt gatgctaaag ggctttcaat agaccttgta agtgaaggag
     6901 atggagccgt caatccgcta ccctgcctct ggtcagttgg tctcagcaat gtaccctgtg
     6961 agtcttgata gactagttgg tgactagtct ctgtagatgg aacaaatggt gcttgagagg
     7021 gcacgtatgg cagagctcca gtcatctgcg gaacatatac tgggcccggg aagatctcat
     7081 ggtcatagct gtttccgctg agggtttaat taagacctca gccgagacag cagaatcacc
     7141 gcccaagtta agcctttgtg ctgatcatgc tctcgaacgg gccaagttcg ggaaaagcaa
     7201 aggagcgttt agtgaggggc aatttgactc acctcccagg caacagatga ggggggcaaa
     7261 aagaaagaaa ttttcgtgag tcaatatgga ttccgagcat cattttcttg cggtctatct
     7321 tgctacgtat gttgatcttg acgctgtgga tcaagcaacg ccactcgctc gctccatcgc
     7381 aggctggtcg cagacaaatt aaaaggcggc aaactcgtac agccgcgggg ttgtccgctg
     7441 caaagtacag agtgataaaa gccgccatgc gaccatcaac gcgttgatgc ccagcttttt
     7501 cgatccgaga atccaccgta gaggcgatag caagtaaaga aaagctaaac aaaaaaaaat
     7561 ttctgcccct aagccatgaa aacgagatgg ggtggagcag aaccaaggaa agagtcgcgc
     7621 tgggctgccg ttccggaagg tgttgtaaag gctcgacgcc caaggtggga gtctaggaga
     7681 agaatttgca tcgggagtgg ggcgggttac ccctccatat ccaatgacag atatctacca
     7741 gccaagggtt tgagcccgcc cgcttagtcg tcgtcctcgc ttgcccctcc ataaaaggat
     7801 ttcccctccc cctcccacaa aattttcttt cccttcctct ccttgtccgc ttcagtacgt
     7861 atatcttccc ttccctcgct tctctcctcc atccttcttt catccatctc ctgctaactt
     7921 ctctgctcag cacctctacg cattactagc cgtagtatct gagcacttct cccttttata
     7981 ttccacaaaa cataacacaa ccttcaccat ggacaagaag tatagcatcg ggctggacat
     8041 tggaacgaac tcggttggtt gggctgtgat tacggacgaa tacaaggtgc catccaagaa
     8101 gtttaaggtc ctgggaaaca ccgaccgtca ctcaatcaag aagaatctca ttggagccct
     8161 gctcttcgat agtggggaga ccgccgaagc tactcgactg aagcgaacgg ctcgccggcg
     8221 ttatacacga cgcaagaatc gcatctgcta cctccaggag attttcagca acgaaatggc
     8281 taaggttgat gactcattct ttcatcgact cgaagaaagt ttcttggtcg aggaggataa
     8341 gaagcacgag cgccatccga tctttggtaa cattgtggat gaggttgcct atcacgaaaa
     8401 gtacccaact atctatcatc ttcgtaagaa gctggtcgat agcacggaca aggctgattt
     8461 gcgacttatc tacctggcac tcgcgcacat gattaagttc cgcggccatt ttcttatcga
     8521 gggtgacctg aaccccgata attctgacgt tgataagctc ttcatccagt tggtccaaac
     8581 ctacaatcag ctgtttgagg aaaaccctat taatgcatct ggcgtggacg ccaaggctat
     8641 cctttcggcg cgcctgtcta agtcgcggcg tttggagaac cttatcgcac aactccccgg
     8701 cgaaaagaag aacggcctct tcggtaattt gattgcgttg tcacttggtc tgactcctaa
     8761 cttcaagagt aattttgacc tggcagagga tgcgaagctc cagttgtcta aggatacgta
     8821 tgatgacgat ctcgacaact tgcttgccca aatcggtgac cagtacgctg atcttttcct
     8881 ggccgctaag aatctctcag atgcaatcct gctcagtgac attttgcggg tcaacaccga
     8941 gattactaag gcccccctgt cagctagtat gatcaagcgg tatgatgagc accatcagga
     9001 cctcaccttg cttaaggccc tcgtgcgtca gcaattgcct gagaagtata aggaaatctt
     9061 ctttgaccaa tccaagaacg gatacgcagg gtatattgat ggcggtgcga gccaggagga
     9121 attctacaag tttatcaagc cgattttgga gaagatggac ggcactgagg aactgctcgt
     9181 caagctgaat cgcgaagatt tgcttcgtaa gcaacgaacg ttcgacaacg gctccatccc
     9241 gcaccagatt catctgggcg agctccacgc catccttcga cgccaggaag atttctaccc
     9301 atttctgaag gacaaccgtg agaagatcga aaagattctt acattccgaa tcccctacta
     9361 tgtgggacct ttggcccgtg ggaattcccg atttgcttgg atgacccgaa agagcgagga
     9421 aaccatcact ccgtggaact tcgaggaagt cgtggacaag ggtgcatccg cgcagagctt
     9481 cattgagcgg atgaccaatt ttgataagaa ccttccgaat gaaaaggtcc tgccaaagca
     9541 ttcgctgctc tacgagtatt tcaccgtgta taacgaactg actaaggtca agtacgtgac
     9601 ggagggaatg cggaagccag ccttcctctc aggggaacaa aagaaggcta tcgtcgattt
     9661 gctttttaag accaatcgta aagtgactgt taagcagctg aaggaggatt atttcaagaa
     9721 gattgaatgt ttcgactccg tcgagatcag cggcgtggaa gatcgcttta acgcttccct
     9781 cggtacctac cacgacctgc tcaagatcat taaggacaag gatttcctcg ataacgagga
     9841 aaatgaggac atcttggaag atattgtcct cacgttgaca ctttttgagg accgcgaaat
     9901 gatcgaggaa cggctcaaga catatgccca tttgttcgac gataaggtga tgaagcagct
     9961 gaagcggcgt cgatacaccg gatggggtcg ccttagccgg aagctgatca acggcattcg
    10021 agataagcaa tctggtaaga ctatcttgga tttccttaag tcggacggct tcgccaaccg
    10081 caattttatg cagcttattc acgacgattc cctgacgttc aaggaggaca tccagaaggc
    10141 acaagtctca ggacaagggg attccctgca cgagcatatc gccaacctgg ctggatcccc
    10201 ggcgatcaag aaggggattc ttcagaccgt caaggttgtc gacgagctgg tcaaggtgat
    10261 gggccgtcat aagccagaaa acatcgtgat tgagatggcc cgagaaaatc agaccactca
    10321 aaagggtcag aagaacagcc gcgagcggat gaagcggatc gaggaaggca ttaaggaact
    10381 tggttctcag atcctgaagg agcaccctgt tgaaaacaca cagctccaaa atgagaagct
    10441 gtatctctac tatttgcaaa atggacgcga catgtacgtc gatcaggagc tcgacattaa
    10501 ccggttgtcg gactacgatg ttgaccatat cgtcccgcaa tccttcctta aggacgatag
    10561 cattgataac aaggtgctga ctcgctcaga taagaaccgg ggcaagtccg acaatgttcc
    10621 aagcgaggaa gtggttaaga agatgaagaa ctactggcgc caattgctta atgccaagct
    10681 catcacacag cgcaagtttg acaacttgac caaggccgag cggggagggc tgagtgaact
    10741 cgataaggct ggcttcatca agcgtcaact cgtggagacg cgacagatca caaagcacgt
    10801 tgctcagatt ctggactccc ggatgaacac aaagtacgac gagaatgata agctcatccg
    10861 tgaagttaag gtcattaccc tcaagtctaa gttggtgtcg gatttccgca aggacttcca
    10921 attttataag gttcgggaga tcaacaatta tcaccatgca catgatgcgt acctcaacgc
    10981 agtcgtggga actgcgctca tcaagaagta tcccaagttg gagtccgaat tcgtctacgg
    11041 ggattataag gtttacgacg tccgcaagat gatcgccaag agtgagcagg aaattggcaa
    11101 ggccacggct aagtatttct tttactccaa catcatgaat ttctttaaga cggagatcac
    11161 actcgccaat ggagaaatcc gtaagcgacc tttgattgag accaacggcg agactggtga
    11221 aatcgtttgg gataaggggc gcgacttcgc taccgtgcgg aaggttctga gcatgccgca
    11281 agtcaatatc gtcaagaaaa ccgaggtgca gacaggcggt ttctctaagg aatcgattct
    11341 tccaaagcgt aactctgaca agctgatcgc tcgaaagaag gattgggacc ccaagaagta
    11401 tggagggttc gattctccta cagtggcata ctcggttctc gttgtcgcga aggttgagaa
    11461 gggaaagtct aagaagctga agtcggtcaa ggaactgctc gggatcacca ttatggagcg
    11521 ctccagcttc gaaaagaatc ccatcgactt tctcgaggcc aagggctata aggaagtcaa
    11581 gaaggatctt atcattaagc tgcctaagta ctctttgttc gagcttgaaa acggtcgaaa
    11641 gcgaatgctc gcatcggcag gagagttgca gaaggggaat gaattggcac ttccctcaaa
    11701 gtacgtgaac ttcctgtatc tcgcgtccca ctacgagaag ctgaagggta gccctgagga
    11761 caacgaacag aagcaacttt ttgttgagca acacaagcat tatctggatg agatcattga
    11821 acagatttca gagttcagta agcgcgtcat cctcgccgat gctaatctcg acaaggtgtt
    11881 gtcggcctac aacaagcacc gtgacaagcc gatccgagag caggctgaaa atatcattca
    11941 tctgttcacc ctcactaact tgggagcacc agcagcgttc aagtattttg atacgacaat
    12001 cgaccgtaag cgatacacgt ccacaaagga ggtgcttgat gcgaccctga ttcatcaatc
    12061 catcactggg ctctatgaaa cccgtatcga ccttagtcaa ctggggggcg accctcccaa
    12121 gaagaagcgc aaggtctgag cggacattcg atttatgccg ttatgacttc cttaaaaaag
    12181 cctttacgaa tgaaagaaat ggaattagac ttgttatgta gttgattcta caatggatta
    12241 tgattcctga acttcaaatc cgctgttcat tattaatctc agctcttccc gtaaagccaa
    12301 tgttgaaact attcgtaaat gtacctcgtt ttgcgtgtac cttgcttatc acgtgatatt
    12361 acatgacctg gacagagttc tgcgcgaaag tcataacgta aatcccgggc ggtaggtgcg
    12421 tcccgggcgg aaggtagttt tctcgtccac cccaacgcgt ttatcaacct caactttcaa
    12481 caaccatcat gccaccaaaa gcgcgtaaaa caaagcgaga tttgattgag caagagggca
    12541 ggatccaatg cgcgattcaa gacattaaaa atggaaaatt tcaaaaaatt gcgcccgcag
    12601 cgcgtgcata caaaattcat cccaatacaa gtgtaatgct agtggaggtc aacacatcaa
    12661 tgcctatttt ggtttagtcg tccaggcggt gagcacaaaa tttgtgtcgt ttgacaagat
    12721 ggttcattta ggcaactggt cagatcagcc ccacttgtag cagtagcggc ggcgctcgaa
    12781 gtgtgactct tattagcaga caggaacgag gacattatta tcatctgctg cttggtgcac
    12841 gataacttgg tgcgtttgtc aagcaaggta agtggacgac ccggtcatac cttcttaagt
    12901 tcgcccttcc tccctttatt tcagattcaa tctgacttac ctattctacc caagcatcca
    12961 aatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
    13021 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
    13081 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
    13141 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
    13201 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
    13261 gcaagacctg cctgaaaccg aactgcccgc tgttctccag ccggtcgcgg aggccatgga
    13321 tgcgattgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
    13381 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
    13441 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
    13501 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcatg cggatttcgg
    13561 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
    13621 gatgttcggg gattcccaat acgaggtcgc caacatcctc ttctggaggc cgtggttggc
    13681 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
    13741 gcgcctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
    13801 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
    13861 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
    13921 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
    13981 ggaatagagt agatgccgac cgatccccac taccgcatta agacctcagc gcggccgcaa
    14041 atttaaataa aatgaagtga agttcctata ctttctagag aataggaact tctatagtga
    14101 gtcgaataag ggcgacacaa aatttattct aaatgcataa taaatactga taacatctta
    14161 tagtttgtat tatattttgt attatcgttg acatgtataa ttttgatatc aaaaactgat
    14221 tttcccttta ttattttcga gatttatttt cttaattctc tttaacaaac tagaaatatt
    14281 gtatatacaa aaaatcataa ataatagatg aatagtttaa ttataggtgt tcatcaatcg
    14341 aaaaagcaac gtatcttatt taaagtgcgt tgcttttttc tcatttataa ggttaaataa
    14401 ttctcatata tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat gctctttccc
    14461 taaactcccc ccataaaaaa acccgccgaa gcgggttttt acgttatttg cggattaacg
    14521 attactcgtt atcagaaccg cccagggggc ccgagcttaa gactggccgt cgttttacaa
    14581 cacagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggggccttc tgcttagttt
    14641 gatgcctggc agttccctac tctcgccttc cgcttcctcg ctcactgact cgctgcgctc
    14701 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
    14761 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
    14821 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
    14881 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
    14941 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
    15001 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
    15061 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
    15121 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
    15181 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
    15241 tgctacagag ttcttgaagt ggtgggctaa ctacggctac actagaagaa cagtatttgg
    15301 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
    15361 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
    15421 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
    15481 cgacgcgcgc gtaactcacg ttaagggatt ttggtcatga gcttgcgccg tcccgtcaag
    15541 tcagcgtaat gctctgctt
//