LOCUS       sequence-165391-       11823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduces DSB at MFE1 locus in Y. lipolytica.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11823)
  AUTHORS   Schwartz C, Shabbir-Hussain M, Frogue K, Blenner M, Wheeldon I
  TITLE     Standardized markerless gene integration for pathway engineering in 
            Yarrowia lipolytica.
  JOURNAL   ACS Synth Biol. 2016 Dec 19.
  PUBMED    27989123
REFERENCE   2  (bases 1 to 11823)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2016 Dec 19."
COMMENT     Sequence Label: pCRISPRyl_MFE1
FEATURES             Location/Qualifiers
     source          1..11823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1740..5843
                     /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             1740..5843
                     /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             5856..5876
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5887..6134
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5903..5921)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6153..6169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6153..6169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6166..6188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6177..6193
                     /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(join(6201..6207,6208..6225,6226..6231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6246..6267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6384..6401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6555..7143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6635..6654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7314..8105,8106..8174))
                     /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     7937..7956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8175..8279)
                     /gene="bla"
                     /label=AmpR promoter
     misc_RNA        8773..8848
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     8898..8916
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccgctttt
      421 cgtagataat ggaatacaaa tggatatcca gagtatacac atggatagta tacactgaca
      481 cgacaattct gtatctcttt atgttaacta ctgtgaggcg ttaaatagag cttgatatat
      541 aaaatgttac atttcacagt ctgaactttt gcagattacc taatttggta agatattaat
      601 tatgaactga aagttgatgg catccctaaa tttgatgaaa gggggatccc ccgggttcga
      661 aggtaccaag gaagcatgcg gtacccgaat tcctgaggtg tctcacaagt gccgtgcagt
      721 cccgccccca cttgcttctc tttgtgtgta gtgtacgtac attatcgaga ccgttgttcc
      781 cgcccacctc gatccggtct agactgaggt gtctcacaag tgccgtgcag tcccgccccc
      841 acttgcttct ctttgtgtgt agtgtacgta cattatcgag accgttgttc ccgcccacct
      901 cgatccgggg atccctgagg tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc
      961 tctttgtgtg tagtgtacgt acattatcga gaccgttgtt cccgcccacc tcgatccggg
     1021 tcgacctgag gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt
     1081 gtagtgtacg tacattatcg agaccgttgt tcccgcccac ctcgatccgg gagctcctga
     1141 ggtgtctcac aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac
     1201 gtacattatc gagaccgttg ttcccgccca cctcgatccg gtctagactg aggtgtctca
     1261 caagtgccgt gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat
     1321 cgagaccgtt gttcccgccc acctcgatcc ggggatccct gaggtgtctc acaagtgccg
     1381 tgcagtcccg cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagaccgt
     1441 tgttcccgcc cacctcgatc cgggtcgacc tgaggtgtct cacaagtgcc gtgcagtccc
     1501 gcccccactt gcttctcttt gtgtgtagtg tacgtacatt atcgagaccg ttgttcccgc
     1561 ccacctcgat ccgggcatgc ctgcagaagc ttttgtggtt gggactttag ccaagggtat
     1621 aaaagaccac cgtccccgaa ttacctttcc tcttcttttc tctctctcct tgtcaactca
     1681 cacccgaaat cgttaagcat ttccttctga gtataagaat cattcaaagg cgcgccacaa
     1741 tggataagaa atactccatt ggcctggaca tcggaaccaa ctccgtgggt tgggccgtga
     1801 tcaccgatga gtacaaggtg ccctctaaga aattcaaggt cctgggcaac accgaccgac
     1861 actccatcaa gaagaacctg atcggcgctc tgctcttcga ctctggcgag accgctgagg
     1921 ccacccgact gaagcgaacc gctcgaagac gatacacccg aagaaagaac cgaatctgtt
     1981 acctgcagga gatcttctct aacgagatgg ccaaggtgga cgactctttc ttccaccgac
     2041 tggaggagtc tttcctggtg gaggaggaca agaagcacga gcgacacccc atcttcggca
     2101 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgcgaaaga
     2161 agctggtgga ctctaccgac aaggccgacc tgcgactgat ctacctggcc ctggcccaca
     2221 tgatcaagtt ccgaggccac ttcctgatcg agggcgacct gaaccccgac aactctgacg
     2281 tggacaagct gttcatccag ctggtgcaga cctacaacca gctcttcgaa gagaacccca
     2341 ttaacgcttc tggcgtggat gctaaggcca tcctgtctgc ccgactgtct aagtctcgac
     2401 gactcgagaa cctgattgct cagctccccg gagagaagaa gaacggtctg ttcggaaacc
     2461 tgattgctct gtccctgggt ctcaccccta acttcaagtc caacttcgat ctggctgagg
     2521 acgctaagct gcagctgtct aaggacacct acgacgatga cctggataac ctgctcgccc
     2581 agattggcga ccagtacgcc gacctgttcc tggccgccaa gaacctgtct gacgccatcc
     2641 tgctgtctga catcctgcga gtgaacaccg agatcaccaa ggcccccctg tctgcctcca
     2701 tgattaagcg atacgatgag caccaccagg atctgaccct cctcaaggct ctggtccgac
     2761 agcagctgcc cgagaagtac aaggagattt tcttcgacca gtctaagaac ggctacgccg
     2821 gctacatcga cggcggcgcc tctcaggagg agttctacaa gttcattaag cccatcctgg
     2881 agaagatgga cggaaccgag gaactgctcg tgaagctgaa ccgagaggac ctcctgcgaa
     2941 agcagcgaac cttcgacaac ggctctatcc cccaccagat ccacctgggc gagctgcacg
     3001 ccatcctgcg acgacaggag gacttctacc ccttcctgaa ggacaaccga gagaagatcg
     3061 agaagatcct gaccttccga atcccctact acgtgggacc cctggcccga ggaaactctc
     3121 gattcgcttg gatgacccga aagtctgagg agaccattac cccctggaac ttcgaggagg
     3181 tggtggataa gggcgcctct gctcagtctt tcatcgagcg aatgaccaac ttcgacaaga
     3241 acctccccaa cgagaaggtc ctgcccaagc actctctgct ctacgagtac ttcaccgtct
     3301 acaacgagct caccaaggtc aagtacgtga ccgagggaat gcgaaagccc gctttcctgt
     3361 ctggagagca gaagaaggct attgtggatc tgctcttcaa gactaaccga aaggtcaccg
     3421 tcaagcagct gaaggaggat tacttcaaga agattgagtg tttcgattct gtcgagatct
     3481 ccggcgtcga ggaccgattc aacgcctctc tgggtaccta ccacgacctg ctgaagatta
     3541 tcaaggacaa ggatttcctg gataacgagg agaacgagga tattctcgag gacattgtcc
     3601 tgaccctcac cctgttcgag gatcgagaga tgattgagga gcgactcaag acctacgctc
     3661 acctgttcga cgacaaggtg atgaagcagc tgaagcgacg acgatacacc ggctggggcc
     3721 gactgtctcg aaagctgatc aacggcatcc gagacaagca gtctggcaag accatcctgg
     3781 acttcctgaa gtctgacggc ttcgccaacc gaaacttcat gcagctgatc cacgacgact
     3841 ctctgacctt caaggaggac atccagaagg cccaggtgtc tggccagggc gactctctgc
     3901 acgagcacat cgccaacctg gccggctctc ccgccattaa gaaaggtatc ctgcagaccg
     3961 tcaaggtggt cgatgagctc gtcaaggtga tgggccgaca caagcccgag aacattgtca
     4021 ttgagatggc tcgagagaac cagactactc agaagggtca gaaaaactcc cgagagcgaa
     4081 tgaagcgaat tgaggaaggt attaaggagc tgggatccca gattctcaag gagcatcccg
     4141 tggagaacac tcagctccag aacgagaagc tgtacctgta ctatctgcag aacggtcgag
     4201 acatgtacgt cgaccaggag ctggatatca accgactctc cgactacgat gtggaccaca
     4261 ttgtgcccca gtccttcctg aaggacgatt ctatcgataa caaggtgctg acccgatccg
     4321 acaagaaccg aggcaagtct gacaacgtgc cctccgagga ggtggtcaag aagatgaaga
     4381 actactggcg acagctgctg aacgccaagc tgattaccca gcgaaagttc gacaacctga
     4441 ccaaggccga gcgaggcggc ctgtctgagc tggacaaggc cggcttcatc aagcgacagc
     4501 tggtggagac ccgacagatc accaagcacg tggcccagat cctggactct cgaatgaaca
     4561 ccaagtacga cgagaacgac aagctgatcc gagaggtgaa ggtgatcacc ctgaagtcta
     4621 agctggtgtc tgacttccga aaggacttcc agttctacaa ggtgcgagag attaacaact
     4681 accaccacgc ccacgatgcc tacctgaacg ctgtcgtggg caccgccctc atcaagaagt
     4741 atcccaagct ggagtccgag ttcgtctacg gcgactacaa ggtctacgat gtgcgaaaaa
     4801 tgattgccaa gtccgagcag gagattggca aggctaccgc caagtacttc ttctactcca
     4861 acattatgaa cttcttcaag accgagatta ccctggctaa cggcgagatt cgaaagcgac
     4921 ccctcattga gaccaacgga gagaccggtg agatcgtgtg ggacaaggga cgagacttcg
     4981 ccaccgtgcg aaaggtgctg tctatgcccc aggtgaacat cgtgaagaag accgaggtgc
     5041 agaccggagg tttctctaag gagtccatcc tgcccaagcg aaactctgac aagctgatcg
     5101 cccgaaagaa ggactgggac cccaagaagt acggaggttt cgactctccc accgtggctt
     5161 actctgtgct ggtggtggcc aaggtggaga agggcaagtc taagaagctg aagtctgtga
     5221 aggagctgct gggcattacc atcatggagc gatcttcttt cgagaagaac cccattgact
     5281 tcctggaggc caagggatac aaggaggtga agaaagatct gattatcaag ctccccaagt
     5341 actctctgtt cgagctggag aacggacgaa agcgaatgct ggcctctgcc ggcgagctgc
     5401 agaagggaaa cgagctggcc ctgccctcca agtacgtcaa cttcctgtac ctcgcctccc
     5461 attacgagaa gctgaagggc tctcccgagg ataacgagca gaagcagctc ttcgtggagc
     5521 agcataagca ctacctggac gagatcatcg agcagatctc tgagttctct aagcgagtga
     5581 tcctggctga cgccaacctg gataaggtgc tgtctgctta caacaagcac cgagacaagc
     5641 ccattcgaga gcaggctgag aacatcattc acctgttcac cctgaccaac ctgggagccc
     5701 ccgctgcctt caagtacttc gacaccacca tcgaccgaaa gcgatacacc tctaccaagg
     5761 aggtgctgga cgccaccctg atccaccagt ctatcaccgg cctgtacgag acccgaatcg
     5821 acctgtctca gctgggcggc gactctcgag ccgaccccaa gaagaagcga aaggtgtaag
     5881 ctagcctcat gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc
     5941 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag
     6001 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac
     6061 gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga
     6121 aggctttaat ttgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6181 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6241 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6301 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6361 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6421 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6481 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6541 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6601 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6661 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6721 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6781 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6841 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6901 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6961 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7021 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7081 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7141 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7201 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7261 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7321 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7381 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7441 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7501 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7561 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7621 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7681 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7741 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7801 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7861 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7921 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7981 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8041 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8101 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8161 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8221 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8281 acatttcccc gaaaagtgcc acctgacgtc cccagttgca aaagttgaca caactctaga
     8341 tctgcttcca aatatagaat cataacaagg gttagggtgt gattatataa tattggtctt
     8401 aattgatgtg ctagggcttt aaaagttggt taaaataacg ctctaatgcc tttttaatat
     8461 attgtctttt tcaaaatctc aaatcggaca cttcttcgtg tatgagactc cattttttgg
     8521 ctccgtcacg tgatatgtat tatcagctat agtggtgtaa acaaagtttt ttactagctg
     8581 taatggcatt ttgtcggagt ggtaaatcgc cttcttgttg tgcgttcgag ttctggactc
     8641 tgcactgggc tactttgaaa aatacctcta atgcgccgat ggtttagtgg taaaatccat
     8701 cgttgccatc gatgggcccc cggttcgatt ccgggtcggc gcaggttgac gtagtctcac
     8761 cgaaaccaac cagttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     8821 acttgaaaaa gtggcaccga gtcggtgctt ttttttacgt ctaagaaacc attattatca
     8881 tgacattaac ctataaaaat aggcgtatca cgaggcccag atcctctaga gtcgaagcgg
     8941 ccgctatgtc tgataaaagg atgtaacata ggcaagctgc tcgtgagtgt tgagtacgaa
     9001 ccttagatcc aaatcacccg cacccacgga tatacttgct tgaatataca gtagtatgcg
     9061 gccgcttcga caccatatca tataaaacta acaatgcatt gaattaatac gaagactacc
     9121 cgttgctatc tccacaccgt tatctccacg gtccaaaggc tgctcaatgt gctgcatacg
     9181 taacgtgggg tgcaaccttg agcacatagt acttttccga aaaccggcga taattaagtg
     9241 tgcactccaa cttttcacac tgagcgtaaa atgtggagaa gaaatcggca ctaaaaagtc
     9301 aggtagactg gaaaatgcgc catgaaatga atatctcttg ctacagtaat gcccagcatc
     9361 gaggggtatt gtgtcaccaa cactatagtg gcagctgaag cgctcgtgat tgtagtatga
     9421 gtctttattg gtgatgggaa gagttcactc aatattctcg ttactgccaa aacaccacgg
     9481 taatcggcca gacaccatgg atgtagatca ccaagcctgt gaatgttatt cgagctaaaa
     9541 tgcacatggt tggtgaaagg agtagttgct gtcgaattcc gtcgtcgcct gagtcatcat
     9601 ttatttacca gttggccaca aacccttgac gatctcgtat gtcccctccg acatactccc
     9661 ggccggctgg ggtacgttcg atagcgctat cggcatcgac aaggtttggg tccctagccg
     9721 ataccgcact acctgagtca caatcttcgg aggtttagtc ttccacatag cacgggcaaa
     9781 agtgcgtata tatacaagag cgtttgccag ccacagattt tcactccaca caccacatca
     9841 cacatacaac cacacacatc cacaatggaa cccgaaacta agaagaccaa gactgactcc
     9901 aagaagattg ttcttctcgg cggcgacttc tgtggccccg aggtgattgc cgaggccgtc
     9961 aaggtgctca agtctgttgc tgaggcctcc ggcaccgagt ttgtgtttga ggaccgactc
    10021 attggaggag ctgccattga gaaggagggc gagcccatca ccgacgctac tctcgacatc
    10081 tgccgaaagg ctgactctat tatgctcggt gctgtcggag gcgctgccaa caccgtatgg
    10141 accactcccg acggacgaac cgacgtgcga cccgagcagg gtctcctcaa gctgcgaaag
    10201 gacctgaacc tgtacgccaa cctgcgaccc tgccagctgc tgtcgcccaa gctcgccgat
    10261 ctctccccca tccgaaacgt tgagggcacc gacttcatca ttgtccgaga gctcgtcgga
    10321 ggtatctact ttggagagcg aaaggaggat gacggatctg gcgtcgcttc cgacaccgag
    10381 acctactccg ttcctgaggt tgagcgaatt gcccgaatgg ccgccttcct ggcccttcag
    10441 cacaaccccc ctcttcccgt gtggtctctt gacaaggcca acgtgctggc ctcctctcga
    10501 ctttggcgaa agactgtcac tcgagtcctc aaggacgaat tcccccagct cgagctcaac
    10561 caccagctga tcgactcggc cgccatgatc ctcatcaagc agccctccaa gatgaatggt
    10621 atcatcatca ccaccaacat gtttggcgat atcatctccg acgaggcctc cgtcatcccc
    10681 ggttctctgg gtctgctgcc ctccgcctct ctggcttctc tgcccgacac caacgaggcg
    10741 ttcggtctgt acgagccctg tcacggatct gcccccgatc tcggcaagca gaaggtcaac
    10801 cccattgcca ccattctgtc tgccgccatg atgctcaagt tctctcttaa catgaagccc
    10861 gccggtgacg ctgttgaggc tgccgtcaag gagtccgtcg aggctggtat cactaccgcc
    10921 gatatcggag gctcttcctc cacctccgag gtcggagact tgttgccaac aaggtcaagg
    10981 agctgctcaa gaaggagtaa gtcgtttcta cgacgcattg atggaaggag caaactgacg
    11041 cgcctgcggg ttggtctacc ggcagggtcc gctagtgtat aagactctat aaaaagggcc
    11101 ctgccctgct aatgaaatga tgatttataa tttaccggtg tagcaacctt gactagaaga
    11161 agcagattgg gtgtgtttgt agtggaggac agtggtacgt tttggaaaca gtcttcttga
    11221 aagtgtcttg tctacagtat attcactcat aacctcaata gccaagggtg tagtcggttt
    11281 attaaaggaa gggagttgtg gctgatgtgg atagatatct ttaagctggc gactgcaccc
    11341 aacgagtgtg gtggtagctt gttactgtat attcggtaag atatattttg tggggtttta
    11401 gtggtgtttg gtaggttagt gcttggtata tgagttgtag gcatgacaat ttggaaaggg
    11461 gtggactttg ggaatattgt gggatttcaa taccttagtt tgtacagggt aattgttaca
    11521 aatgatacaa agaactgtat ttcttttcat ttgttttaat tggttgtata tcaagtccgt
    11581 tagacgagct cagtgccatg gcttttggca ctgtatttca tttttagagg tacactacat
    11641 tcagtgaggt atggtaaggt tgagggcata atgaaggcac cttgtactga cagtcacaga
    11701 cctctcaccg agaattttat gagatatact cgggttcatt ttaggctccg attcgattca
    11761 aattattact gtcgaaatcg gttgagcatc cgttgatttc cgaacagatc tggccctttc
    11821 gtc
//
