LOCUS       sequence-256284-       12721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LYS5ex_CrisprCas9-yl_RFP: CAS9 vector with LYS5ex marker for gRNA 
            cloning using GoldenGate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12721)
  AUTHORS   Larroude M, Trabelsi H, Nicaud JM, Rossignol T
  TITLE     A set of Yarrowia lipolytica CRISPR/Cas9 vectors for exploiting 
            wild-type strain diversity.
  JOURNAL   Biotechnol Lett. 2020 Jan 23. pii: 10.1007/s10529-020-02805-4. doi: 
            10.1007/s10529-020-02805-4.
  PUBMED    31974649
REFERENCE   2  (bases 1 to 12721)
  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: "Biotechnol 
            Lett. 2020 Jan 23. pii: 10.1007/s10529-020-02805-4. doi: 
            10.1007/s10529-020-02805-4."
COMMENT     Sequence Label: JME4393
FEATURES             Location/Qualifiers
     source          1..12721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      12..69
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(93..110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(264..852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(344..363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1023..1814,1815..1883))
                     /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     1646..1665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1884..1988)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2078..2121)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     protein_bind    2700..2721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2736..2741,2742..2759,2760..2766)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2774..2790
                     /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)."
     RBS             2808..2819
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2826..3503
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3548..3619
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3635..3662
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    complement(4007..4040)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(6334..6356)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6456..6475
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6669..6691
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6683..6700
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6684..6700
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8313..12416
                     /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             8313..12416
                     /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             12429..12449
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      12460..12707
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(12476..12494)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 tcttcctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa
       61 ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc
      121 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      181 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      241 aaaaaggccg cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa
      301 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      361 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      421 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      481 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      541 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      601 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      661 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      721 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      781 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      841 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
      901 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
      961 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1021 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1081 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1141 cccagtgctg caatgatacc gcgcgaccca cgctcaccgg ctccagattt atcagcaata
     1201 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1261 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1321 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1381 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1441 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1501 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1561 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1621 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1681 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1741 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     1801 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1861 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     1921 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     1981 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     2041 acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta
     2101 gctttcgcta aggatgattt ctggaattcg aagagcctct gacctgacgt ccccagttgc
     2161 aaaagttgac acaactctag atctgcttcc aaatatagaa tcataacaag ggttagggtg
     2221 tgattatata atattggtct taattgatgt gctagggctt taaaagttgg ttaaaataac
     2281 gctctaatgc ctttttaata tattgtcttt ttcaaaatct caaatcggac acttcttcgt
     2341 gtatgagact ccattttttg gctccgtcac gtgatatgta ttatcagcta tagtggtgta
     2401 aacaaagttt tttactagct gtaatggcat tttgtcggag tggtaaatcg ccttcttgtt
     2461 gtgcgttcga gttctggact ctgcactggg ctactttgaa aaatacctct aatgcgccga
     2521 tggtttagtg gtaaaatcca tcgttgccat cgatgggccc ccggttcgag agacggggcc
     2581 ggccatctgg gccctagagc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2641 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2701 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     2761 atgttgtgtg gaattgtgag cggataacaa tttcacacat actagagaaa gaggagaaat
     2821 actagatggc ttcctccgaa gacgttatca aagagttcat gcgtttcaaa gttcgtatgg
     2881 aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg
     2941 aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg
     3001 acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac ccggctgaca
     3061 tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt gttatgaact
     3121 tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac ggtgagttca
     3181 tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt atgcagaaaa
     3241 aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt gctctgaaag
     3301 gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct gaagttaaaa
     3361 ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac
     3421 tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa cgtgctgaag
     3481 gtcgtcactc caccggtgct taataacgct gatagtgcta gtgtagatcg ctactagagc
     3541 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     3601 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     3661 cgtttatata ggccatctgg gccgggggcg tctctgagct agaaatagca agttaaaata
     3721 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tttacgtcta
     3781 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggcccagatc
     3841 ctctagagtc gaagcggccg ctatgtctga taaaaggatg taacataggc aagctgctcg
     3901 tgagtgttga gtacgaacct tagatccaaa tcacccgcac ccacggatat acttgcttga
     3961 atatacagta gtatgcggcc gctaggtatt accctgttat ccctacataa cttcgtatag
     4021 catacatcat acgaagttat tctgaattcc gggtggaaat gtgagtattg acacaaggat
     4081 cgccacggac gcctgtctga tacgagcggc ctattacagg aacgtgcatt gatgatacga
     4141 gcaagtgtac gtgaatcgac cgatgcacag ccataaggag ccacttctgc aactactggc
     4201 tcagctactt atgctgctac tactaattaa ctgggactgc ttccgctaac caagtgattt
     4261 ttttatggtg gcggcctagt agcgtgatta gtagtgcggt tgtctagtac agtgcaggtg
     4321 gaagtgcgga agttgcacat gtaagtgcaa gtggacttga agggggtgtt ttttttagtg
     4381 ccttggcttg tttttgcgac cgcctcattg ccagactcac atacacacac gcattgctct
     4441 gggtgttgga acggttcttg ccacccaatt gcgccatgaa ggtgtaaatg ggtatgggca
     4501 gatgactcag ggtttttctg tcttacagct gaagtttacc cccccacccc aacgcgtgcg
     4561 tgtgatattt tttgtgatat ctctccccag ggttggcctt acaatttcaa tccgcccctc
     4621 gcccccactt ccacagtgtg tctttcacac gcaccgagct ttgtctgttt ttttttttgt
     4681 ttttctctcc aactgccaac atctgtctta ccatcgttgt tgtttgcatg cgtggttggt
     4741 gtttttttgg ttttgatttt gcatttttcg ttttcatcaa ggacataaat gttcacctcg
     4801 tgtgccaaag tacacttttc acataacgat ggtgactgcc agacttgtag cgacgacatg
     4861 gccaatattc tgtggcgatg ggggttgtgc tctgcgtttc tcctgctctc ctcggagttt
     4921 tgttcctgat gtacgtccac acaccttcag ccttcaatac taacacagct ttttgattgg
     4981 caaaactcca agcaatccgt ctatttgttt ctgctggacg tgcggcccat ctggaactcc
     5041 cagaacgact tcaaccaggc catgcgactg cttcctgtcg aatctcagct cgacatttcg
     5101 cgttttcggt acagaagcga tgcgaagatg gctctcggct ctcagctgct ccagcggtat
     5161 attgccaccg ttctgacagg caagaagtgg acggacattg tcatttcgaa gactccgcag
     5221 ggcaagtcat ttgtggccaa ctcgccgttc gactacaaca tcagtcacca cagcgagctg
     5281 gtggttgtgg ctgttcgtcc tgatggcaag cagattggag tggatatcac ctcgactgtt
     5341 cctccctcaa actggactat gaactggatg gagggctttt cacagatttt ctcgccgttc
     5401 gagtactctc aggtcaccag cgacagcacc ggcaatggcg agttcttctt gcggtgggca
     5461 ctcaaggagt cgtattccaa ggccattggc atggggctca acatggacct tgccaagcta
     5521 gagttcagaa acctggacac ggccagactg atggacgcgg ccaaggacaa gaccggaacc
     5581 gacgttgtgg gcgccgtggc tgtccagaga ggctggagca actctgcgca gctgtggatt
     5641 gatggagtcg cacaggacag atggcaccac gagattttca tgctcaacaa caatcatgtg
     5701 atcgctctgg cgacacacaa ggagggtgtg aatgaggtgg agcaggtgcg actgagaaat
     5761 ttaacggtgg aggagttgat gagcagagcg ttgccttttg gaccggctgt ggccatgcag
     5821 cctcattcca acccggtcta atttgcacgt ctagaatttg aacctatcgg cagcacataa
     5881 acacacgtgc acacacacac acacacacac acacacacac acacacatac tcagccagac
     5941 ttgcgcgacg tcgtacacag ctggaggcca agtcctcaca acacacggtc tgcatcaact
     6001 gagtccgctc tgaagagtgg aacatcatcc gtactgtagg agcacggttc atgcctctgc
     6061 aatgtagata gatacatata attgaacagc atgtaatata cactattgga agcagtggat
     6121 gaataaaacc tatgtagcga cggtatggta atggtaatgg cacacccacc aggtgtcttc
     6181 aagtctgctt atttgccgga attcgtataa cttcgtatag caggagttat ccgaagcgat
     6241 aattaccctg ttatccctag atcgattccc acggatccgg gatatcctaa cggggccctt
     6301 tcgtcacgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggtgac
     6361 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6421 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     6481 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6541 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     6601 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     6661 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacccg
     6721 cttttcgtag ataatggaat acaaatggat atccagagta tacacatgga tagtatacac
     6781 tgacacgaca attctgtatc tctttatgtt aactactgtg aggcgttaaa tagagcttga
     6841 tatataaaat gttacatttc acagtctgaa cttttgcaga ttacctaatt tggtaagata
     6901 ttaattatga actgaaagtt gatggcatcc ctaaatttga tgaaaggggg aacccccggg
     6961 ttcgaaggta ccaaggaagc atgcggtacc cgaattcgct tcgatacgcg tatcgatgat
     7021 acgcgtccat ggtgaggtgt ctcacaagtg ccgtgcagtc ccgcccccac ttgcttctct
     7081 ttgtgtgtag tgtacgtaca ttatcgagag ggttgttccc gcccacctcg atccggcatg
     7141 ctgaggtgtc tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt tgtgtgtagt
     7201 gtacgtacat tatcgagagg gttgttcccg cccacctcga tccggcatgc tgaggtgtct
     7261 cacaagtgcc gtgcagtccc gcccccactt gcttctcttt gtgtgtagtg tacgtacatt
     7321 atcgagaggg ttgttcccgc ccacctcgat ccggcatgct gaggtgtctc acaagtgccg
     7381 tgcagtcccg cccccacttg cttctctttg tgtgtagtgt acgtacatta tcgagagggt
     7441 tgttcccgcc cacctcgatc cggcatgctg aggtgtctca caagtgccgt gcagtcccgc
     7501 ccccacttgc ttctctttgt gtgtagtgta cgtacattat cgagagggtt gttcccgccc
     7561 acctcgatcc ggcatgctga ggtgtctcac aagtgccgtg cagtcccgcc cccacttgct
     7621 tctctttgtg tgtagtgtac gtacattatc gagagggttg ttcccgccca cctcgatccg
     7681 gcatgctgag gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt
     7741 gtagtgtacg tacattatcg agagggttgt tcccgcccac ctcgatccga catgttgagg
     7801 tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc tctttgtgtg tagtgtacgt
     7861 acattatcga gagggttgtt cccgcccacc tcgatccggc atgcactgat cacgggcaaa
     7921 agtgcttcga tagagagggg gttggcggcg catttgtgtc ccaaaaaaca gccccaattg
     7981 ccccaattga ccccaaattg acccagtagc ggacccaacc ccggcgagag cccccttcac
     8041 cccacatatc aaacctcccc cggttcccac acttgccgtt aagggcgtag ggtactgcag
     8101 tctggaatct acgcttgttc agactttgta ctagtttctt tgtctggcca tccgggtaac
     8161 ccatgccgga cgcaaaatag actactgaaa atttttttgc tttgtggttg ggactttagc
     8221 caagggtata aaagaccacc gtccccgaat tacctttcct cttcttttct ctctctcctt
     8281 gtcaactcac acccgaagga tccacaacca caatggataa gaaatactcc attggcctgg
     8341 acatcggaac caactccgtg ggttgggccg tgatcaccga tgagtacaag gtgccctcta
     8401 agaaattcaa ggtcctgggc aacaccgacc gacactccat caagaagaac ctgatcggcg
     8461 ctctgctctt cgactctggc gagactgctg aggccacccg actgaagcga accgctcgaa
     8521 gacgatacac ccgaagaaag aaccgaatct gttacctgca ggagatcttc tctaacgaga
     8581 tggccaaggt ggacgactct ttcttccacc gactggagga gtctttcctg gtggaggagg
     8641 acaagaagca cgagcgacac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     8701 agaagtaccc caccatctac cacctgcgaa agaagctggt ggactctacc gacaaggccg
     8761 acctgcgact gatctacctg gccctggccc acatgatcaa gttccgaggc cacttcctga
     8821 tcgagggcga cctgaacccc gacaactctg acgtggacaa gctgttcatc cagctggtgc
     8881 agacctacaa ccagctcttc gaagagaacc ccattaacgc ttctggcgtg gatgctaagg
     8941 ccatcctgtc tgcccgactg tctaagtctc gacgactcga gaacctgatt gctcagctcc
     9001 ccggagagaa gaagaacggt ctgttcggaa acctgattgc tctgtccctg ggtctgaccc
     9061 ctaacttcaa gtccaacttc gatctggctg aggacgctaa gctgcagctg tctaaggaca
     9121 cctacgacga tgacctggat aacctgctcg cccagattgg cgaccagtac gccgacctgt
     9181 tcctggccgc caagaacctg tctgacgcca tcctgctgtc tgacatcctg cgagtgaaca
     9241 ccgagatcac caaggccccc ctgtctgcct ccatgattaa gcgatacgat gagcaccacc
     9301 aggatctgac cctcctcaag gctctggtcc gacagcagct gcccgagaag tacaaggaga
     9361 ttttcttcga ccagtctaag aacggctacg ccggctacat cgacggcggc gcctctcagg
     9421 aggagttcta caagttcatt aagcccatcc tggagaagat ggacggaacc gaggaactgc
     9481 tcgtgaagct gaaccgagag gacctcctgc gaaagcagcg aaccttcgac aacggctcta
     9541 tcccccacca gatccacctg ggcgagctgc acgccatcct gcgacgacag gaggacttct
     9601 accccttcct gaaggacaac cgagagaaga tcgagaagat cctgaccttc cgaatcccct
     9661 actacgtggg acccctggcc cgaggaaact ctcgattcgc ttggatgacc cgaaagtctg
     9721 aggagactat taccccctgg aacttcgagg aggtggtgga taagggcgcc tctgctcagt
     9781 ctttcatcga gcgaatgacc aacttcgaca agaacctccc caacgagaag gtcctgccca
     9841 agcactctct gctctacgag tacttcaccg tctacaacga gctcaccaag gtcaagtacg
     9901 tgaccgaggg aatgcgaaag cccgctttcc tgtctggaga gcagaagaag gctattgtgg
     9961 atctgctctt caagactaac cgaaaggtca ccgtcaagca gctgaaggag gattacttca
    10021 agaagattga gtgtttcgat tctgtcgaga tctccggcgt cgaggaccga ttcaacgcct
    10081 ctctgggtac ctaccacgac ctgctgaaga ttatcaagga caaggatttc ctggataacg
    10141 aggagaacga ggatattctc gaggacattg tcctgaccct caccctgttc gaggatcgag
    10201 agatgattga ggagcgactc aagacctacg ctcacctgtt cgacgacaag gtgatgaagc
    10261 agctgaagcg acgacgatac accggctggg gccgactgtc tcgaaagctg atcaacggca
    10321 tccgagacaa gcagtctggc aagaccatcc tggacttcct gaagtctgac ggcttcgcca
    10381 accgaaactt catgcagctg atccacgacg actctctgac cttcaaggag gacatccaga
    10441 aggcccaggt gtctggccag ggcgactctc tgcacgagca catcgccaac ctggccggct
    10501 ctcccgccat taagaaaggt atcctgcaga ccgtcaaggt ggtcgatgag ctcgtcaagg
    10561 tgatgggccg acacaagccc gagaacattg tcattgagat ggctcgagag aaccagacta
    10621 ctcagaaggg tcagaaaaac tcccgagagc gaatgaagcg aattgaggaa ggtattaagg
    10681 agctgggctc ccagattctc aaggagcatc ccgtggagaa cactcagctc cagaacgaga
    10741 agctgtacct gtactatctg cagaacggtc gagacatgta cgtcgaccag gagctggata
    10801 tcaaccgact ctccgactac gatgtggacc acattgtgcc ccagtccttc ctgaaggacg
    10861 attctatcga taacaaggtg ctgacccgat ccgacaagaa ccgaggcaag tctgacaacg
    10921 tgccctccga ggaggtggtc aagaagatga agaactactg gcgacagctg ctgaacgcca
    10981 agctgattac ccagcgaaag ttcgacaacc tgaccaaggc cgagcgaggc ggcctgtctg
    11041 agctggacaa ggccggcttc atcaagcgac agctggtgga gactcgacag atcaccaagc
    11101 acgtggccca gatcctggac tctcgaatga acaccaagta cgacgagaac gacaagctga
    11161 tccgagaggt gaaggtgatc accctgaagt ctaagctggt gtctgacttc cgaaaggact
    11221 tccagttcta caaggtgcga gagattaaca actaccacca cgcccacgat gcctacctga
    11281 acgctgtcgt gggcaccgcc ctcatcaaga agtatcccaa gctggagtcc gagttcgtct
    11341 acggcgacta caaggtctac gatgtgcgaa aaatgattgc caagtccgag caggagattg
    11401 gcaaggctac cgccaagtac ttcttctact ccaacattat gaacttcttc aagaccgaga
    11461 ttaccctggc taacggcgag attcgaaagc gacccctcat tgagactaac ggagagactg
    11521 gtgagatcgt gtgggacaag ggacgagact tcgccaccgt gcgaaaggtg ctgtctatgc
    11581 cccaggtgaa catcgtgaag aagaccgagg tgcagaccgg aggtttctct aaggagtcca
    11641 tcctgcccaa gcgaaactct gacaagctga tcgcccgaaa gaaggactgg gaccccaaga
    11701 agtacggagg tttcgactct cccaccgtgg cttactctgt gctggtggtg gccaaggtgg
    11761 agaagggcaa gtctaagaag ctgaagtctg tgaaggagct gctgggcatt accatcatgg
    11821 agcgatcttc tttcgagaag aaccccattg acttcctgga ggccaaggga tacaaggagg
    11881 tgaagaaaga tctgattatc aagctcccca agtactctct gttcgagctg gagaacggac
    11941 gaaagcgaat gctggcctct gccggcgagc tgcagaaggg aaacgagctg gccctgccct
    12001 ccaagtacgt caacttcctg tacctcgcct cccattacga gaagctgaag ggctctcccg
    12061 aggataacga gcagaagcag ctcttcgtgg agcagcataa gcactacctg gacgagatca
    12121 tcgagcagat ttctgagttc tctaagcgag tgatcctggc tgacgccaac ctggataagg
    12181 tgctgtctgc ttacaacaag caccgagaca agcccattcg agagcaggct gagaacatca
    12241 ttcacctgtt caccctgacc aacctgggag cccccgctgc cttcaagtac ttcgacacca
    12301 ccatcgaccg aaagcgatac acctctacca aggaggtgct ggacgccacc ctgatccacc
    12361 agtctatcac cggcctgtac gagactcgaa tcgacctgtc tcagctgggc ggcgactctc
    12421 gagccgaccc caagaagaag cgaaaggtgt aagctagcct catgtaatta gttatgtcac
    12481 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
    12541 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
    12601 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
    12661 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag cttggcggtc
    12721 t
//
