pCRISPRyl

LOCUS       pCRISPRyl               11805 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  CRISPR/Cas9 vector for Yarrowia lipolytica, with AvrII site for 
            sgRNA insertion.
ACCESSION   .
VERSION     .
KEYWORDS    pCRISPRyl
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11805)
  AUTHORS   Schwartz CM, Hussain MS, Blenner M, Wheeldon I
  TITLE     Synthetic RNA polymerase III promoters facilitate high efficiency 
            CRISPR-Cas9 mediated genome editing in Yarrowia lipolytica.
  JOURNAL   ACS Synth Biol. 2015 Dec 29.
  PUBMED    26714206
REFERENCE   2  (bases 1 to 11805)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11805
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(2908..2926)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_RNA        complement(2976..3051)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3528..3631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3632..4492
                     /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     complement(3850..3869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4663..5251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5152..5171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5405..5422
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5539..5560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5575..5605
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5613..5629
                     /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)."
     primer_bind     5618..5640
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5637..5653
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5637..5653
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      5672..5919
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5885..5903
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(5930..5950)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5963..10066)
                     /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"
     primer_bind     complement(11411..11428)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11411..11427)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11420..11442)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(11636..11655)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11755..11777
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg ccagatctgt tcggaaatca acggatgctc aaccgatttc gacagtaata
       61 atttgaatcg aatcggagcc taaaatgaac ccgagtatat ctcataaaat tctcggtgag
      121 aggtctgtga ctgtcagtac aaggtgcctt cattatgccc tcaaccttac catacctcac
      181 tggatgtagt gtacctctaa aaatgaaata cagtgccaaa agccatggca ctgagctcgt
      241 ctaacggact tgatatacaa ccaattaaaa caaatgaaaa gaaatacagt tctttgtatc
      301 atttgtaaca attaccctgt acaaactaag gtattgaaat cccacaatat tcccaaagtc
      361 cacccctttc caaattgtca tgcctacaac tcatatacca gacactaacc taccaaacac
      421 cactaaaaac cccacaaaat atatcttacc gaatatacag taacaagcta ccaccacact
      481 cgttgggtgc agtcgccagc tttaaagata tctatccaca tcagccacaa ctcccttcct
      541 ttaataaacc gactacaccc ttggctattg aggttatgag tgaatatact gtagacaaga
      601 cactttcaag aagactgttt ccaaaacgta ccactgtcct ccactacaaa cacacccaat
      661 ctgcttcttc tagtcaaggt tgctacaccg gtaaattata aatcatcatt tcattagcag
      721 ggctgggccc tttttataga gtcttataca ctagcggacc ctgccggtag accaacccgc
      781 aggcgcgtca gtttgctcct tccatcaatg cgtcgtagaa acgacttact ccttcttgag
      841 cagctccttg accttgttgg caacaaagtc tccgacctcg gaggtggagg aagagcctcc
      901 gatatcggcg gtagtgatac cagcctcgac ggactccttg acggcagcct caacagcgtc
      961 accggcgggc ttcatgttaa gagagaactt gagcatcatg gcggcagaca gaatggtggc
     1021 aatggggttg accttctgct tgccgagatc gggggcagat ccgtgacagg gctcgtacag
     1081 accgaacgcc tcgttggtgt cgggcagaga agccagagag gcggagggca gcagacccag
     1141 agaaccgggg atgacggagg cctcgtcgga gatgatatcg ccaaacatgt tggtggtgat
     1201 gatgatacca ttcatcttgg agggctgctt gatgaggatc atggcggccg agtcgatcag
     1261 ctggtggttg agctcgagct gggggaattc gtccttgagg actcgagtga cagtctttcg
     1321 ccaaagtcga gaggaggcca gcacgttggc cttgtcaaga gaccacacgg gaagaggggg
     1381 gttgtgctga agggccagga aggcggccat tcgggcaatt cgctcaacct caggaacgga
     1441 gtaggtctcg gtgtcggaag cgacgccaga tccgtcatcc tcctttcgct ctccaaagta
     1501 gatacctccg acgagctctc ggacaatgat gaagtcggtg ccctcaacgt ttcggatggg
     1561 ggagagatcg gcgagcttgg gcgacagcag ctggcagggt cgcaggttgg cgtacaggtt
     1621 caggtccttt cgcagcttga ggagaccctg ctcgggtcgc acgtcggttc gtccgtcggg
     1681 agtggtccat acggtgttgg cagcgcctcc gacagcaccg agcataatag agtcagcctt
     1741 tcggcagatg tcgagagtag cgtcggtgat gggctcgccc tccttctcaa tggcagctcc
     1801 tccaatgagt cggtcctcaa acacaaactc ggtgccggag gcctcagcaa cagacttgag
     1861 caccttgacg gcctcggcaa tcacctcggg gccacagaag tcgccgccga gaagaacaat
     1921 cttcttggag tcagtcttgg tcttcttagt ttcgggttcc attgtggatg tgtgtggttg
     1981 tatgtgtgat gtggtgtgtg gagtgaaaat ctgtggctgg caaacgctct tgtatatata
     2041 cgcacttttg cccgtgctat gtggaagact aaacctccga agattgtgac tcaggtagtg
     2101 cggtatcggc tagggaccca aaccttgtcg atgccgatag cgctatcgaa cgtaccccag
     2161 ccggccggga gtatgtcgga ggggacatac gagatcgtca agggtttgtg gccaactggt
     2221 aaataaatga tgactcaggc gacgacggaa ttcgacagca actactcctt tcaccaacca
     2281 tgtgcatttt agctcgaata acattcacag gcttggtgat ctacatccat ggtgtctggc
     2341 cgattaccgt ggtgttttgg cagtaacgag aatattgagt gaactcttcc catcaccaat
     2401 aaagactcat actacaatca cgagcgcttc agctgccact atagtgttgg tgacacaata
     2461 cccctcgatg ctgggcatta ctgtagcaag agatattatt tcatggcgca ttttccagtc
     2521 tacctgactt tttagtgtga tttcttctcc acattttatg ctcagtgtga aaagttggag
     2581 tgcacactta attatcgccg gttttcggaa agtactatgt gctcaaggtt gcaccccacg
     2641 ttacgtatgc agcacattga gcagcctttg gaccgtggag ataacggtgt ggagatagca
     2701 acgggtagtc ttcgtattaa ttcaatgcat tgttagtttt atatgatatg gtgtcgaagc
     2761 ggccgcatac tactgtatat tcaagcaagt atatccgtgg gtgcgggtga tttggatcta
     2821 aggttcgtac tcaacactca cgagcagctt gcctatgtta catcctttta tcagacatag
     2881 cggccgcttc gactctagag gatctgggcc tcgtgatacg cctattttta taggttaatg
     2941 tcatgataat aatggtttct tagacgtaaa aaaaagcacc gactcggtgc cactttttca
     3001 agttgataac ggactagcct tattttaact tgctatttct agctctaaaa cctaggtcaa
     3061 cctgcgccga cccggaatcg aaccgggggc ccatcgatgg caacgatgga ttttaccact
     3121 aaaccatcgg cgcattagag gtatttttca aagtagccca gtgcagagtc cagaactcga
     3181 acgcacaaca agaaggcgat ttaccactcc gacaaaatgc cattacagct agtaaaaaac
     3241 tttgtttaca ccactatagc tgataataca tatcacgtga cggagccaaa aaatggagtc
     3301 tcatacacga agaagtgtcc gatttgagat tttgaaaaag acaatatatt aaaaaggcat
     3361 tagagcgtta ttttaaccaa cttttaaagc cctagcacat caattaagac caatattata
     3421 taatcacacc ctaacccttg ttatgattct atatttggaa gcagatctag agttgtgtca
     3481 acttttgcaa ctggggacgt caggtggcac ttttcgggga aatgtgtgcg gaacccctat
     3541 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3601 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3661 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3721 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3781 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3841 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3901 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3961 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4021 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4081 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4141 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4201 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4261 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4321 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4381 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4441 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4501 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4561 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4621 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4681 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4741 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4801 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4861 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4921 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4981 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5041 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5101 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5161 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5221 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5281 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5341 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5401 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     5461 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5521 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     5581 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     5641 aaacagctat gaccatgatt acgccaagct tgcaaattaa agccttcgag cgtcccaaaa
     5701 ccttctcaag caaggttttc agtataatgt tacatgcgta cacgcgtctg tacagaaaaa
     5761 aaagaaaaat ttgaaatata aataacgttc ttaatactaa cataactata aaaaaataaa
     5821 tagggaccta gacttcaggt tgtctaactc cttccttttc ggttagagcg gatgtggagg
     5881 gagggcgtga atgtaagcgt gacataacta attacatgag gctagcttac acctttcgct
     5941 tcttcttggg gtcggctcga gagtcgccgc ccagctgaga caggtcgatt cgggtctcgt
     6001 acaggccggt gatagactgg tggatcaggg tggcgtccag cacctccttg gtagaggtgt
     6061 atcgctttcg gtcgatggtg gtgtcgaagt acttgaaggc agcgggggct cccaggttgg
     6121 tcagggtgaa caggtgaatg atgttctcag cctgctctcg aatgggcttg tctcggtgct
     6181 tgttgtaagc agacagcacc ttatccaggt tggcgtcagc caggatcact cgcttagaga
     6241 actcagagat ctgctcgatg atctcgtcca ggtagtgctt atgctgctcc acgaagagct
     6301 gcttctgctc gttatcctcg ggagagccct tcagcttctc gtaatgggag gcgaggtaca
     6361 ggaagttgac gtacttggag ggcagggcca gctcgtttcc cttctgcagc tcgccggcag
     6421 aggccagcat tcgctttcgt ccgttctcca gctcgaacag agagtacttg gggagcttga
     6481 taatcagatc tttcttcacc tccttgtatc ccttggcctc caggaagtca atggggttct
     6541 tctcgaaaga agatcgctcc atgatggtaa tgcccagcag ctccttcaca gacttcagct
     6601 tcttagactt gcccttctcc accttggcca ccaccagcac agagtaagcc acggtgggag
     6661 agtcgaaacc tccgtacttc ttggggtccc agtccttctt tcgggcgatc agcttgtcag
     6721 agtttcgctt gggcaggatg gactccttag agaaacctcc ggtctgcacc tcggtcttct
     6781 tcacgatgtt cacctggggc atagacagca cctttcgcac ggtggcgaag tctcgtccct
     6841 tgtcccacac gatctcaccg gtctctccgt tggtctcaat gaggggtcgc tttcgaatct
     6901 cgccgttagc cagggtaatc tcggtcttga agaagttcat aatgttggag tagaagaagt
     6961 acttggcggt agccttgcca atctcctgct cggacttggc aatcattttt cgcacatcgt
     7021 agaccttgta gtcgccgtag acgaactcgg actccagctt gggatacttc ttgatgaggg
     7081 cggtgcccac gacagcgttc aggtaggcat cgtgggcgtg gtggtagttg ttaatctctc
     7141 gcaccttgta gaactggaag tcctttcgga agtcagacac cagcttagac ttcagggtga
     7201 tcaccttcac ctctcggatc agcttgtcgt tctcgtcgta cttggtgttc attcgagagt
     7261 ccaggatctg ggccacgtgc ttggtgatct gtcgggtctc caccagctgt cgcttgatga
     7321 agccggcctt gtccagctca gacaggccgc ctcgctcggc cttggtcagg ttgtcgaact
     7381 ttcgctgggt aatcagcttg gcgttcagca gctgtcgcca gtagttcttc atcttcttga
     7441 ccacctcctc ggagggcacg ttgtcagact tgcctcggtt cttgtcggat cgggtcagca
     7501 ccttgttatc gatagaatcg tccttcagga aggactgggg cacaatgtgg tccacatcgt
     7561 agtcggagag tcggttgata tccagctcct ggtcgacgta catgtctcga ccgttctgca
     7621 gatagtacag gtacagcttc tcgttctgga gctgagtgtt ctccacggga tgctccttga
     7681 gaatctggga tcccagctcc ttaatacctt cctcaattcg cttcattcgc tctcgggagt
     7741 ttttctgacc cttctgagta gtctggttct ctcgagccat ctcaatgaca atgttctcgg
     7801 gcttgtgtcg gcccatcacc ttgacgagct catcgaccac cttgacggtc tgcaggatac
     7861 ctttcttaat ggcgggagag ccggccaggt tggcgatgtg ctcgtgcaga gagtcgccct
     7921 ggccagacac ctgggccttc tggatgtcct ccttgaaggt cagagagtcg tcgtggatca
     7981 gctgcatgaa gtttcggttg gcgaagccgt cagacttcag gaagtccagg atggtcttgc
     8041 cagactgctt gtctcggatg ccgttgatca gctttcgaga cagtcggccc cagccggtgt
     8101 atcgtcgtcg cttcagctgc ttcatcacct tgtcgtcgaa caggtgagcg taggtcttga
     8161 gtcgctcctc aatcatctct cgatcctcga acagggtgag ggtcaggaca atgtcctcga
     8221 gaatatcctc gttctcctcg ttatccagga aatccttgtc cttgataatc ttcagcaggt
     8281 cgtggtaggt acccagagag gcgttgaatc ggtcctcgac gccggagatc tcgacagaat
     8341 cgaaacactc aatcttcttg aagtaatcct ccttcagctg cttgacggtg acctttcggt
     8401 tagtcttgaa gagcagatcc acaatagcct tcttctgctc tccagacagg aaagcgggct
     8461 ttcgcattcc ctcggtcacg tacttgacct tggtgagctc gttgtagacg gtgaagtact
     8521 cgtagagcag agagtgcttg ggcaggacct tctcgttggg gaggttcttg tcgaagttgg
     8581 tcattcgctc gatgaaagac tgagcagagg cgcccttatc caccacctcc tcgaagttcc
     8641 agggggtaat ggtctcctca gactttcggg tcatccaagc gaatcgagag tttcctcggg
     8701 ccaggggtcc cacgtagtag gggattcgga aggtcaggat cttctcgatc ttctctcggt
     8761 tgtccttcag gaaggggtag aagtcctcct gtcgtcgcag gatggcgtgc agctcgccca
     8821 ggtggatctg gtgggggata gagccgttgt cgaaggttcg ctgctttcgc aggaggtcct
     8881 ctcggttcag cttcacgagc agttcctcgg ttccgtccat cttctccagg atgggcttaa
     8941 tgaacttgta gaactcctcc tgagaggcgc cgccgtcgat gtagccggcg tagccgttct
     9001 tagactggtc gaagaaaatc tccttgtact tctcgggcag ctgctgtcgg accagagcct
     9061 tgaggagggt cagatcctgg tggtgctcat cgtatcgctt aatcatggag gcagacaggg
     9121 gggccttggt gatctcggtg ttcactcgca ggatgtcaga cagcaggatg gcgtcagaca
     9181 ggttcttggc ggccaggaac aggtcggcgt actggtcgcc aatctgggcg agcaggttat
     9241 ccaggtcatc gtcgtaggtg tccttagaca gctgcagctt agcgtcctca gccagatcga
     9301 agttggactt gaagttaggg gtgagaccca gggacagagc aatcaggttt ccgaacagac
     9361 cgttcttctt ctctccgggg agctgagcaa tcaggttctc gagtcgtcga gacttagaca
     9421 gtcgggcaga caggatggcc ttagcatcca cgccagaagc gttaatgggg ttctcttcga
     9481 agagctggtt gtaggtctgc accagctgga tgaacagctt gtccacgtca gagttgtcgg
     9541 ggttcaggtc gccctcgatc aggaagtggc ctcggaactt gatcatgtgg gccagggcca
     9601 ggtagatcag tcgcaggtcg gccttgtcgg tagagtccac cagcttcttt cgcaggtggt
     9661 agatggtggg gtacttctcg tggtaggcca cctcgtccac gatgttgccg aagatggggt
     9721 gtcgctcgtg cttcttgtcc tcctccacca ggaaagactc ctccagtcgg tggaagaaag
     9781 agtcgtccac cttggccatc tcgttagaga agatctcctg caggtaacag attcggttct
     9841 ttcttcgggt gtatcgtctt cgagcggttc gcttcagtcg ggtggcctca gcggtctcgc
     9901 cagagtcgaa gagcagagcg ccgatcaggt tcttcttgat ggagtgtcgg tcggtgttgc
     9961 ccaggacctt gaatttctta gagggcacct tgtactcatc ggtgatcacg gcccaaccca
    10021 cggagttggt tccgatgtcc aggccaatgg agtatttctt atccattgtg gcgcgccttt
    10081 gaatgattct tatactcaga aggaaatgct taacgatttc gggtgtgagt tgacaaggag
    10141 agagagaaaa gaagaggaaa ggtaattcgg ggacggtggt cttttatacc cttggctaaa
    10201 gtcccaacca caaaagcttc tgcaggcatg cccggatcga ggtgggcggg aacaacggtc
    10261 tcgataatgt acgtacacta cacacaaaga gaagcaagtg ggggcgggac tgcacggcac
    10321 ttgtgagaca cctcaggtcg acccggatcg aggtgggcgg gaacaacggt ctcgataatg
    10381 tacgtacact acacacaaag agaagcaagt gggggcggga ctgcacggca cttgtgagac
    10441 acctcaggga tccccggatc gaggtgggcg ggaacaacgg tctcgataat gtacgtacac
    10501 tacacacaaa gagaagcaag tgggggcggg actgcacggc acttgtgaga cacctcagtc
    10561 tagaccggat cgaggtgggc gggaacaacg gtctcgataa tgtacgtaca ctacacacaa
    10621 agagaagcaa gtgggggcgg gactgcacgg cacttgtgag acacctcagg agctcccgga
    10681 tcgaggtggg cgggaacaac ggtctcgata atgtacgtac actacacaca aagagaagca
    10741 agtgggggcg ggactgcacg gcacttgtga gacacctcag gtcgacccgg atcgaggtgg
    10801 gcgggaacaa cggtctcgat aatgtacgta cactacacac aaagagaagc aagtgggggc
    10861 gggactgcac ggcacttgtg agacacctca gggatccccg gatcgaggtg ggcgggaaca
    10921 acggtctcga taatgtacgt acactacaca caaagagaag caagtggggg cgggactgca
    10981 cggcacttgt gagacacctc agtctagacc ggatcgaggt gggcgggaac aacggtctcg
    11041 ataatgtacg tacactacac acaaagagaa gcaagtgggg gcgggactgc acggcacttg
    11101 tgagacacct caggaattcg ggtaccgcat gcttccttgg taccttcgaa cccgggggat
    11161 ccccctttca tcaaatttag ggatgccatc aactttcagt tcataattaa tatcttacca
    11221 aattaggtaa tctgcaaaag ttcagactgt gaaatgtaac attttatata tcaagctcta
    11281 tttaacgcct cacagtagtt aacataaaga gatacagaat tgtcgtgtca gtgtatacta
    11341 tccatgtgta tactctggat atccatttgt attccattat ctacgaaaag cgggtaccga
    11401 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
    11461 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
    11521 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
    11581 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
    11641 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
    11701 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
    11761 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//