LOCUS       sequence-190010-        7653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows inducible expression of gRNAs and Cas9 for plasmid curing..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 7110 to 7144)
  AUTHORS   Lauritsen I, Porse A, Sommer MOA, Norholm MHH
  TITLE     A versatile one-step CRISPR-Cas9 based approach to plasmid-curing.
  JOURNAL   Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 
            10.1186/s12934-017-0748-z.
  PUBMED    28764701
REFERENCE   2  (bases 1 to 7653)
  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: "Microb Cell
            Fact."; date: "2017-08-2"; volume: "16(1):135. doi"; pages: " 
            10.1186/s12934-017-0748-z"
COMMENT     Sequence Label: pFREE_Cm
FEATURES             Location/Qualifiers
     source          1..7653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             130..4236
                     /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             130..4233
                     /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(4271..4289)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4285..4332
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             4466..5089
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(4539..4558)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      complement(5207..5301)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5322..5981)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISKWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5896..5915
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5982..6084)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      complement(6147..6782)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(6292..6311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_RNA        complement(7031..7109)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     promoter        complement(7110..7144)
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     repeat_region   7281..7316
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   7347..7382
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   7413..7448
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   7479..7514
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   7545..7580
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacagaagg
       61 agatatacat ggataagaaa tactgagcac tatcagtgat agagatactg agcacagaag
      121 gagatataca tggataagaa atactcaata ggcttagata tcggcacaaa tagcgtcgga
      181 tgggcggtga tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat
      241 acagaccgcc acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag
      301 acagcggaag cgactcgtct caaacggaca gctcgtagaa ggtatacacg tcggaagaat
      361 cgtatttgtt atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc
      421 tttcatcgac ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct
      481 atttttggaa atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat
      541 ctgcgaaaaa aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc
      601 ttagcgcata tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat
      661 aatagtgatg tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa
      721 gaaaacccta ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt
      781 aaatcaagac gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta
      841 tttgggaatc tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat
      901 ttggcagaag atgctaaatt acagctttca aaagatactt acgatgatga tttagataat
      961 ttattggcgc aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca
     1021 gatgctattt tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta
     1081 tcagcttcaa tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct
     1141 ttagttcgac aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac
     1201 ggatatgcag gttatattga tgggggagct agccaagaag aattttataa atttatcaaa
     1261 ccaattttag aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat
     1321 ttgctgcgca agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt
     1381 gagctgcatg ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt
     1441 gagaagattg aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt
     1501 ggcaatagtc gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat
     1561 tttgaagaag ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac
     1621 tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat
     1681 tttacggttt ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca
     1741 gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga
     1801 aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt
     1861 gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg
     1921 ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag
     1981 gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa
     2041 acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact
     2101 ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa
     2161 acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc
     2221 catgatgata gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc
     2281 gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt
     2341 ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa
     2401 aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg
     2461 cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa
     2521 gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa
     2581 aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat
     2641 gtcgatcaca ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta
     2701 acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa
     2761 aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca acgtaagttt
     2821 gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc
     2881 aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt
     2941 cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc
     3001 ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag
     3061 attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg
     3121 attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat
     3181 gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc
     3241 ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt
     3301 cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg
     3361 cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa
     3421 acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac
     3481 aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca
     3541 acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta
     3601 aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat
     3661 ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa
     3721 ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc
     3781 ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat
     3841 ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg
     3901 tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag tgaattttct
     3961 aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat
     4021 agagacaaac caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat
     4081 cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg
     4141 tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa
     4201 acacgcattg atttgagtca gctaggaggt gactgagatc ccatgttacc ggtatcctag
     4261 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4321 aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta
     4381 gtcaataaac cggtattgac gccaacgggt cacacgggta cagtgctagc agaaacatat
     4441 ccatgaaatc ccgccccgaa ttcatatgtc tagattagat aaaagtaaag tgattaacag
     4501 cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca
     4561 gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct
     4621 cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg
     4681 ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag
     4741 tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac
     4801 tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata
     4861 tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca
     4921 agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca
     4981 agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt
     5041 gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtcttaag caatagacat
     5101 aagcggctat ttaacgaccc tgccctgaac cgacgacaag ctgacgaccg ggtctccgca
     5161 agtggcactt ttcggggaaa tgtgcgcgga acccctatgt gcttggattc tcaccaataa
     5221 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     5281 ctggatctat caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac
     5341 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     5401 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     5461 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     5521 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     5581 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     5641 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     5701 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     5761 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     5821 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     5881 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccatttg
     5941 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     6001 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     6061 ttggaacctc ttacgtgccg atcaacgtct cattttcgcc agatatcgac gtctaagaaa
     6121 ccatacaaat aggcatgcta gcgcagaaac gtcctagaag atgccaggag gatacttagc
     6181 agagagacaa taaggccgga gcgaagccgt ttttccatag gctccgcccc cctgacgaac
     6241 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     6301 aggcgtttcc ccctgatggc tccctcttgc gctctcctgt tcccgtcctg cggcgtccgt
     6361 gttgtggtgg aggctttacc caaatcacca cgtcccgttc cgtgtagaca gttcgctcca
     6421 agctgggctg tgtgcaagaa ccccccgttc agcccgactg ctgcgcctta tccggtaact
     6481 atcatcttga gtccaacccg gaaagacacg acaaaacgcc actggcagca gccattggta
     6541 actgagaatt agtggattta gatatcgaga gtcttgaagt ggtggcctaa cagaggctac
     6601 actgaaagga cagtatttgg tatctgcgct ccactaaagc cagttaccag gttaagcagt
     6661 tccccaactg acttaacctt cgatcaaacc gcctccccag gcggtttttt cgtttacaga
     6721 gcaggagatt acgacgatcg taaaaggatc tcaagaagat cctttacgga ttcccgacac
     6781 catcactcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
     6841 tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac
     6901 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     6961 tacattaatt gcgttgcgca caccgactag cgaaaaaacc ccgccgaagc ggggtttttt
     7021 gcgaaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt
     7081 taacttgcta tgctgttttg aatggttccg ctagcactgt acctaggact gagctagccg
     7141 tcaaaagcag cgtatatacc atggccacaa ttcagcaaat tgtgaacatc atcacgttca
     7201 tctttccctg gttgccaatg gcccattttc ctgtcagtaa cgagaaggtc gcgaattcag
     7261 gcgcttttta gactggtcgt gttttagagc tatgctgttt tgaatggtcc caaaacatga
     7321 actagcgatt agtcgctatg acttaagttt tagagctatg ctgttttgaa tggtcccaaa
     7381 acaaccacac tagagaacat actggctaaa tagttttaga gctatgctgt tttgaatggt
     7441 cccaaaacgg ttggactcaa gacgatagtt accggatagt tttagagcta tgctgttttg
     7501 aatggtccca aaacggcgaa acccgacagg actataaaga taccgtttta gagctatgct
     7561 gttttgaatg gtcccaaaac aaaaaaaaac cccgcccctg acagggcggg gttttttttt
     7621 taattaaagt ctggtcctcg agtctggtta taa
//
