LOCUS       sequence-381918-        8372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses deactivated-Cas9 (J23107) and tracrRNA, with BBR1 origin, 
            ampR, Start-Stop assembly site for array insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8372)
  TITLE     CRISPRi with randomised arrays
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8372)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pRAC07A
FEATURES             Location/Qualifiers
     source          1..8372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      471..565
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    614..635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(650..655,656..673,674..680)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    688..704
                     /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     712..728
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     712..728
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             724..1047
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    741..797
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(798..815)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(798..814)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(807..829)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1023..1042)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(1075..1161)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        1220..1283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1284..1352,1353..2144)
                     /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(1502..1521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            2439..2547
                     /note="incP origin of transfer"
     CDS             complement(2558..3220)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      3221..3992
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(4107..4193)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_RNA        complement(4398..4476)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     promoter        4547..4581
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     RBS             4588..4599
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(4606..4632,4633..4635,4636..7122,7123..7125,7126..8372,
                     1..340)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDDIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(4606..4632,4633..4635,4636..8372,1..340)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDDIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 attgtttgtg gagcagcata agcattattt agatgagatt attgagcaaa tcagtgaatt
       61 ttctaagcgt gttattttag cagatgccaa tttagataaa gttcttagtg catataacaa
      121 acatagagac aaaccaatac gtgaacaagc agaaaatatt attcatttat ttacgttgac
      181 gaatcttgga gctcccgctg cttttaaata ttttgataca acaattgatc gtaaacgata
      241 tacgtctaca aaagaagttt tagatgccac tcttatccat caatccatca ctggtcttta
      301 tgaaacacgc attgatttga gtcagctagg aggtgactga taagaattgc tcggtaccaa
      361 attccagaaa agagacgctt tcgagcgtct tttttcgttt tggtcctgac gcttacctct
      421 taagaggtca ctgacctaac atttacggct agctcagtcc tagagtcgag gactcctgtt
      481 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
      541 ccgggcgttt tttattggtg agaatggtct caaggtcaga gaagagccag tagggcagtg
      601 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      661 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      721 gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg
      781 gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
      841 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
      901 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
      961 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     1021 agtacaatct gctctgatgc cgcatagttt ccgctcttca ggatactaga gaccatttgt
     1081 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     1141 ctgagccttt cgttttattt gatgcaggca tgcaagcttg cggccgcgtc gtgactggga
     1201 aaaccctggc gactagaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1261 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1321 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1381 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1441 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1501 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1561 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1621 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     1681 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     1741 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     1801 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     1861 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     1921 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     1981 ctggagccgg tgagcgtggg tcccgcggta tcattgcagc actggggcca gatggtaagc
     2041 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2101 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccatgg gcaaatatta
     2161 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     2221 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     2281 cggggcgtaa tttgactttt gtccttttcc gctgcataac cctgcttcgg ggtcattata
     2341 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     2401 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     2461 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     2521 gggaatcctg ctctgcgagg ctggccgtag gccggcccta ccggcgcggc agcgttaccc
     2581 gtgtcggcgg ctccaacggc tcgccatcgt ccagaaaaca cggctcatcg ggcatcggca
     2641 ggcgctgctg cccgcgccgt tcccattcct ccgtttcggt caaggctggc aggtctggtt
     2701 ccatgcccgg aatgccgggc tggctgggcg gctcctcgcc ggggccggtc ggtagttgct
     2761 gctcgcccgg atacagggtc gggatgcggc gcaggtcgcc atgccccaac agcgattcgt
     2821 cctggtcgtc gtgatcaacc accacggcgg cactgaacac cgacaggcgc aactggtcgc
     2881 ggggctggcc ccacgccacg cggtcattga ccacgtaggc cgacacggtg ccggggccgt
     2941 tgagcttcac gacggagatc cagcgctcgg ccaccaagtc cttgactgcg tattggaccg
     3001 tccgcaaaga acgtccgatg agcttggaaa gtgtcttctg gctgaccacc acggcgttct
     3061 ggtggcccat ctgcgccacg aggtgatgca gcagcattgc cgccgtgggt ttcctcgcaa
     3121 taagcccggc ccacgcctca tgcgctttgc gttccgtttg cacccagtga ccgggcttgt
     3181 tcttggcttg aatgccgatt tctctggact gcgtggccat gcttatctcc atgcggtagg
     3241 ggtgccgcac ggttgcggca ccatgcgcaa tcagctgcaa cttttcggca gcgcgacaac
     3301 aattatgcgt tgcgtaaaag tggcagtcaa ttacagattt tctttaacct acgcaatgag
     3361 ctattgcggg gggtgccgca atgagctgtt gcgtaccccc cttttttaag ttgttgattt
     3421 ttaagtcttt cgcatttcgc cctatatcta gttctttggt gcccaaagaa gggcacccct
     3481 gcggggttcc cccacgcctt cggcgcggct ccccctccgg caaaaagtgg cccctccggg
     3541 gcttgttgat cgactgcgcg gccttcggcc ttgcccaagg tggcgctgcc cccttggaac
     3601 ccccgcactc gccgccgtga ggctcggggg gcaggcgggc gggcttcgcc cttcgactgc
     3661 ccccactcgc ataggcttgg gtcgttccag gcgcgtcaag gccaagccgc tgcgcggtcg
     3721 ctgcgcgagc cttgacccgc cttccacttg gtgtccaacc ggcaagcgaa gcgcgcaggc
     3781 cgcaggccgg aggcttttcc ccagagaaaa ttaaaaaaat tgatggggca aggccgcagg
     3841 ccgcgcagtt ggagccggtg ggtatgtggt cgaaggctgg gtagccggtg ggcaatccct
     3901 gtggtcaagc tcgtgggcag gcgcagcctg tccatcagct tgtccagcag ggttgtccac
     3961 gggccgagcg aagcgagcca gccggtggcc gctcgcggcc atcgtccaca tatccacggg
     4021 ctggcaaggg agcgcagcga ccgcgcaggg cgaagcccgg agagcaagcc cgtagggggg
     4081 gcgcgcccag ctgtctaggg cggcggattt gtcctactca ggagagcgtt caccgacaaa
     4141 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctt
     4201 taattaaagc ggataacaat ttcacacagg aggccgccta ggccgcggcc gcgcgaattc
     4261 aaaagatccg gcagcgccta aatgtagaat gataaaagga ttaagagatt aatttcccta
     4321 aaaatgataa aacaagcgtt ttgaaagcgc ttgttttttt ggtttgcagt cagagtagaa
     4381 tagaagtatc aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc
     4441 cttattttaa cttgctatgc tgttttgaat ggttccaaca agattatttt ataactttta
     4501 taacaaataa tcaaggagaa attcaaagaa atttatcagc cactagttta cggctagctc
     4561 agccctaggt attatgctag cagatctaaa gaggagaaag gatctatgga taagaaatac
     4621 tcaataggct tagctatcgg cacaaatagc gtcggatggg cggtgatcac tgatgaatat
     4681 aaggttccgt ctaaaaagtt caaggttctg ggaaatacag accgccacag tatcaaaaaa
     4741 aatcttatag gggctctttt atttgacagt ggagagacag cggaagcgac tcgtctcaaa
     4801 cggacagctc gtagaaggta tacacgtcgg aagaatcgta tttgttatct acaggagatt
     4861 ttttcaaatg agatggcgaa agtagatgat agtttctttc atcgacttga agagtctttt
     4921 ttggtggaag aagacaagaa gcatgaacgt catcctattt ttggaaatat agtagatgaa
     4981 gttgcttatc atgagaaata tccaactatc tatcatctgc gaaaaaaatt ggtagattct
     5041 actgataaag cggatttgcg cttaatctat ttggccttag cgcatatgat taagtttcgt
     5101 ggtcattttt tgattgaggg agatttaaat cctgataata gtgatgtgga caaactattt
     5161 atccagttgg tacaaaccta caatcaatta tttgaagaaa accctattaa cgcaagtgga
     5221 gtagatgcta aagcgattct ttctgcacga ttgagtaaat caagacgatt agaaaatctc
     5281 attgctcagc tccccggtga gaagaaaaat ggcttatttg ggaatctcat tgctttgtca
     5341 ttgggtttga cccctaattt taaatcaaat tttgatttgg cagaagatgc taaattacag
     5401 ctttcaaaag atacttacga tgatgattta gataatttat tggcgcaaat tggagatcaa
     5461 tatgctgatt tgtttttggc agctaagaat ttatcagatg ctattttact ttcagatatc
     5521 ctaagagtaa atactgaaat aactaaggct cccctatcag cttcaatgat taaacgctac
     5581 gatgaacatc atcaagactt gactctttta aaagctttag ttcgacaaca acttccagaa
     5641 aagtataaag aaatcttttt tgatcaatca aaaaacggat atgcaggtta tattgatggg
     5701 ggagctagcc aagaagaatt ttataaattt atcaaaccaa ttttagaaaa aatggatggt
     5761 actgaggaat tattggtgaa actaaatcgt gaagatttgc tgcgcaagca acggaccttt
     5821 gacaacggct ctattcccca tcaaattcac ttgggtgagc tgcatgctat tttgagaaga
     5881 caagaagact tttatccatt tttaaaagac aatcgtgaga agattgaaaa aatcttgact
     5941 tttcgtattc cttattatgt tggtccattg gcgcgtggca atagtcgttt tgcatggatg
     6001 actcggaagt ctgaagaaac aattacccca tggaattttg aagaagttgt cgataaaggt
     6061 gcttcagctc aatcatttat tgaacgcatg acaaactttg ataaaaatct tccaaatgaa
     6121 aaagtactac caaaacatag tttgctttat gagtatttta cggtttataa cgaattgaca
     6181 aaggtcaaat atgttactga aggaatgcga aaaccagcat ttctttcagg tgaacagaag
     6241 aaagccattg ttgatttact cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa
     6301 gaagattatt tcaaaaaaat agaatgtttt gatagtgttg aaatttcagg agttgaagat
     6361 agatttaatg cttcattagg tacctaccat gatttgctaa aaattattaa agataaagat
     6421 tttttggata atgaagaaaa tgaagatatc ttagaggata ttgttttaac attgacctta
     6481 tttgaagata gggagatgat tgaggaaaga cttaaaacat atgctcacct ctttgatgat
     6541 aaggtgatga aacagcttaa acgtcgccgt tatactggtt ggggacgttt gtctcgaaaa
     6601 ttgattaatg gtattaggga taagcaatct ggcaaaacaa tattagattt tttgaaatca
     6661 gatggttttg ccaatcgcaa ttttatgcag ctgatccatg atgatagttt gacatttaaa
     6721 gaagacattc aaaaagcaca agtgtctgga caaggcgata gtttacatga acatattgca
     6781 aatttagctg gtagccctgc tattaaaaaa ggtattttac agactgtaaa agttgttgat
     6841 gaattggtca aagtaatggg gcggcataag ccagaaaata tcgttattga aatggcacgt
     6901 gaaaatcaga caactcaaaa gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa
     6961 gaaggtatca aagaattagg aagtcagatt cttaaagagc atcctgttga aaatactcaa
     7021 ttgcaaaatg aaaagctcta tctctattat ctccaaaatg gaagagacat gtatgtggac
     7081 caagaattag atattaatcg tttaagtgat tatgatgtcg atgacattgt tccacaaagt
     7141 ttccttaaag acgattcaat agacaataag gtcttaacgc gttctgataa aaatcgtggt
     7201 aaatcggata acgttccaag tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa
     7261 cttctaaacg ccaagttaat cactcaacgt aagtttgata atttaacgaa agctgaacgt
     7321 ggaggtttga gtgaacttga taaagctggt tttatcaaac gccaattggt tgaaactcgc
     7381 caaatcacta agcatgtggc acaaattttg gatagtcgca tgaatactaa atacgatgaa
     7441 aatgataaac ttattcgaga ggttaaagtg attaccttaa aatctaaatt agtttctgac
     7501 ttccgaaaag atttccaatt ctataaagta cgtgagatta acaattacca tcatgcccat
     7561 gatgcgtatc taaatgccgt cgttggaact gctttgatta agaaatatcc aaaacttgaa
     7621 tcggagtttg tctatggtga ttataaagtt tatgatgttc gtaaaatgat tgctaagtct
     7681 gagcaagaaa taggcaaagc aaccgcaaaa tatttctttt actctaatat catgaacttc
     7741 ttcaaaacag aaattacact tgcaaatgga gagattcgca aacgccctct aatcgaaact
     7801 aatggggaaa ctggagaaat tgtctgggat aaagggcgag attttgccac agtgcgcaaa
     7861 gtattgtcca tgccccaagt caatattgtc aagaaaacag aagtacagac aggcggattc
     7921 tccaaggagt caattttacc aaaaagaaat tcggacaagc ttattgctcg taaaaaagac
     7981 tgggatccaa aaaaatatgg tggttttgat agtccaacgg tagcttattc agtcctagtg
     8041 gttgctaagg tggaaaaagg gaaatcgaag aagttaaaat ccgttaaaga gttactaggg
     8101 atcacaatta tggaaagaag ttcctttgaa aaaaatccga ttgacttttt agaagctaaa
     8161 ggatataagg aagttaaaaa agacttaatc attaaactac ctaaatatag tctttttgag
     8221 ttagaaaacg gtcgtaaacg gatgctggct agtgccggag aattacaaaa aggaaatgag
     8281 ctggctctgc caagcaaata tgtgaatttt ttatatttag ctagtcatta tgaaaagttg
     8341 aagggtagtc cagaagataa cgaacaaaaa ca
//
