LOCUS       sequence-201645-        7864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified from pEM-Cas9HF1 (Addgene ID: 89961) to co-express 
            inducible recA56 to block recA-mediated double-strand break repair..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7864)
  AUTHORS   Moreb EA, Hoover B, Yaseen A, Valyasevi N, Roecker Z, Menacho-Melgar
            R, Lynch MD
  TITLE     Managing the SOS Response for Enhanced CRISPR-Cas-Based 
            Recombineering in E. coli through Transient Inhibition of Host RecA 
            Activity.
  JOURNAL   ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174.
  PUBMED    28915012
REFERENCE   2  (bases 1 to 7864)
  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. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174."
COMMENT     Sequence Label: pEM-Cas9HF1-indRecA56
FEATURES             Location/Qualifiers
     source          1..7864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      603..674
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      690..717
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(745..762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(879..1424)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(959..978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1538..1632)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1653..2312)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2227..2246
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2313..2415)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(2445..3071)
                     /codon_start=1
                     /gene="tetR"
                     /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="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     2976..2995
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(3084..3118)
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     promoter        3119..3174
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    3155..3173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(3210..4697,4698..4700,4701..5189,5190..5192,5193..5291,
                     5292..5294,5295..5984,5985..5987,5988..7313)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Kleinstiver et al., 2016)"
                     /label=SpCas9-HF1
                     /note="carries the mutations N497A, R661A, Q695A, and 
                     Q926A"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3210..7313
                     /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
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(7376..7864,1..573)
                     /codon_start=1
                     /gene="recA"
                     /product="DNA repair protein from E. coli"
                     /label=RecA
                     /translation="MAIDENKQKALAAALGQIEKQFGKGSIMRLGEDRSMDVETISTGS
                     LSLDIALGAGGLPMGCIVEIYGPESSGKTTLTLQVIAAAQREGKTCAFIDAEHALDPIY
                     ARKLGVDIDNLLCSQPDTGEQALEICDALARSGAVDVIVVDSVAALTPKAEIEGEIGDS
                     HMGLAARMMSQAMRKLAGNLKQSNTLLIFINQIRMKIGVMFGNPETTTGGNALKFYASV
                     RLDIRRIGAVKEGENVVGSETRVKVVKNKIAAPFKQAEFQILYGEGINFYGELVDLGVK
                     EKLIEKAGAWYSYKGEKIGQGKANATAWLKDNPETAKEIEKKVRELLLSNPNSTPDFSV
                     DDSEGVAETNEDF"
ORIGIN
        1 cacatgggcc ttgcggcacg tatgatgagc caggcgatgc gtaagctggc gggtaacctg
       61 aagcagtcca acacgctgct gatcttcatc aaccagatcc gtatgaaaat tggtgtgatg
      121 ttcggtaacc cggaaaccac taccggtggt aacgcgctga aattctacgc ctctgttcgt
      181 ctggacatcc gtcgtatcgg cgcggtgaaa gagggcgaaa acgtggtggg tagcgaaacc
      241 cgcgtgaaag tggtgaagaa caaaatcgct gcgccgttta aacaggctga attccagatc
      301 ctctacggcg aaggtatcaa cttctacggc gaactggttg acctgggcgt aaaagagaag
      361 ctgatcgaga aagcaggcgc gtggtacagc tacaaaggtg agaagatcgg tcagggtaaa
      421 gcgaatgcga ctgcctggct gaaagataac ccggaaaccg cgaaagagat cgagaagaaa
      481 gtacgtgagt tgctgctgag caacccgaac tcaacgccgg atttctctgt agatgatagc
      541 gaaggcgtag cagaaactaa cgaagatttt taagcttaac tcgagtaagg atctccaggc
      601 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      661 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
      721 atacctaggg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
      781 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
      841 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct
      901 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
      961 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     1021 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     1081 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     1141 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     1201 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     1261 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     1321 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     1381 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     1441 atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     1501 tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc
     1561 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     1621 tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag
     1681 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     1741 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     1801 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     1861 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     1921 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     1981 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     2041 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc
     2101 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     2161 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     2221 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     2281 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     2341 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     2401 cttacgtgcc gatcaacgtc tcattttcgc cagatatcga cgtcttaaga cccactttca
     2461 catttaagtt gtttttctaa tccgcatatg atcaattcaa ggccgaataa gaaggctggc
     2521 tctgcacctt ggtgatcaaa taattcgata gcttgtcgta ataatggcgg catactatca
     2581 gtagtaggtg tttccctttc ttctttagcg acttgatgct cttgatcttc caatacgcaa
     2641 cctaaagtaa aatgccccac agcgctgagt gcatataatg cattctctag tgaaaaacct
     2701 tgttggcata aaaaggctaa ttgattttcg agagtttcat actgtttttc tgtaggccgt
     2761 gtacctaaat gtacttttgc tccatcgcga tgacttagta aagcacatct aaaactttta
     2821 gcgttattac gtaaaaaatc ttgccagctt tccccttcta aagggcaaaa gtgagtatgg
     2881 tgcctatcta acatctcaat ggctaaggcg tcgagcaaag cccgcttatt ttttacatgc
     2941 caatacaatg taggctgctc tacacctagc ttctgggcga gtttacgggt tgttaaacct
     3001 tcgattccga cctcattaag cagctctaat gcgctgttaa tcactttact tttatctaat
     3061 ctagacatca ttaattccta attgctagca ttgtacctag gactgagcta gccataaagt
     3121 tgacactcta tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat
     3181 tcaaaagatc taaagaggac ttcggatcta tggataagaa atactcaata ggcttagata
     3241 tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt ccgtctaaaa
     3301 agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc
     3361 ttttatttga cagtggagag acagcggaag cgactcgcct caaacggaca gctcgtagaa
     3421 ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca aatgagatgg
     3481 cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg gaagaagaca
     3541 agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct tatcatgaga
     3601 aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat aaagcggatt
     3661 tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat tttttgattg
     3721 agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag ttggtacaaa
     3781 cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat gctaaagcga
     3841 ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct cagctccccg
     3901 gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt ttgaccccta
     3961 attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca aaagatactt
     4021 acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct gatttgtttt
     4081 tggcagctaa gaatttatca gatgctattt tactttcaga tatcctaaga gtaaatactg
     4141 aaataactaa ggctccccta tcagcttcaa tgattaaacg ctacgatgaa catcatcaag
     4201 acttgactct tttaaaagct ttagttcgac aacaacttcc agaaaagtat aaagaaatct
     4261 tttttgatca atcaaaaaac ggatatgcag gttatattga tgggggagct agccaagaag
     4321 aattttataa atttatcaaa ccaattttag aaaaaatgga tggtactgag gaattattgg
     4381 tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc
     4441 cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc
     4501 catttttaaa agacaatcgt gagaagattg aaaaaatctt gacttttcga attccttatt
     4561 atgttggtcc attggcgcgt ggcaatagtc gttttgcatg gatgactcgg aagtctgaag
     4621 aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat
     4681 ttattgaacg catgacagcg tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac
     4741 atagtttgct ttatgagtat tttacggttt ataacgaatt gacaaaggtc aaatatgtta
     4801 ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatt
     4861 tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa
     4921 aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcat
     4981 taggtaccta ccatgatttg ctaaaaatta ttaaagataa agattttttg gataatgaag
     5041 aaaatgaaga tatcttagag gatattgttt taacattgac cttatttgaa gatagggaga
     5101 tgattgagga aagacttaaa acatatgctc acctctttga tgataaggtg atgaaacagc
     5161 ttaaacgtcg ccgttatact ggttggggag cgttgtctcg aaaattgatt aatggtatta
     5221 gggataagca atctggcaaa acaatattag attttttgaa atcagatggt tttgccaatc
     5281 gcaattttat ggcgctgatc catgatgata gtttgacatt taaagaagac attcaaaaag
     5341 cacaagtgtc tggacaaggc gatagtttac atgaacatat tgcaaattta gctggtagcc
     5401 ctgctattaa aaaaggtatt ttacagactg taaaagttgt tgatgaattg gtcaaagtaa
     5461 tggggcggca taagccagaa aatatcgtta ttgaaatggc acgtgaaaat cagacaactc
     5521 aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat
     5581 taggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc
     5641 tctatctcta ttatctccaa aatggaagag acatgtatgt ggaccaagaa ttagatatta
     5701 atcgtttaag tgattatgat gtcgatcaca ttgttccaca aagtttcctt aaagacgatt
     5761 caatagacaa taaggtctta acgcgttctg ataaaaatcg tggtaaatcg gataacgttc
     5821 caagtgaaga agtagtcaaa aagatgaaaa actattggag acaacttcta aacgccaagt
     5881 taatcactca acgtaagttt gataatttaa cgaaagctga acgtggaggt ttgagtgaac
     5941 ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgcgcgatc actaagcatg
     6001 tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttattc
     6061 gagaggttaa agtgattacc ttaaaatcta aattagtttc tgacttccga aaagatttcc
     6121 aattctataa agtacgtgag attaacaatt accatcatgc ccatgatgcg tatctaaatg
     6181 ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaatcggag tttgtctatg
     6241 gtgattataa agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca
     6301 aagcaaccgc aaaatatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta
     6361 cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga aactaatggg gaaactggag
     6421 aaattgtctg ggataaaggg cgagattttg ccacagtgcg caaagtattg tccatgcccc
     6481 aagtcaatat tgtcaagaaa acagaagtac agacaggcgg attctccaag gagtcaattt
     6541 taccaaaaag aaattcggac aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat
     6601 atggtggttt tgatagtcca acggtagctt attcagtcct agtggttgct aaggtggaaa
     6661 aagggaaatc gaagaagtta aaatccgtta aagagttact agggatcaca attatggaaa
     6721 gaagttcctt tgaaaaaaat ccgattgact ttttagaagc taaaggatat aaggaagtta
     6781 aaaaagactt aatcattaaa ctacctaaat atagtctttt tgagttagaa aacggtcgta
     6841 aacggatgct ggctagtgcc ggagaattac aaaaaggaaa tgagctggct ctgccaagca
     6901 aatatgtgaa ttttttatat ttagctagtc attatgaaaa gttgaagggt agtccagaag
     6961 ataacgaaca aaaacaattg tttgtggagc agcataagca ttatttagat gagattattg
     7021 agcaaatcag tgaattttct aagcgtgtta ttttagcaga tgccaattta gataaagttc
     7081 ttagtgcata taacaaacat agagacaaac caatacgtga acaagcagaa aatattattc
     7141 atttatttac gttgacgaat cttggagctc ccgctgcttt taaatatttt gatacaacaa
     7201 ttgatcgtaa acgatatacg tctacaaaag aagttttaga tgccactctt atccatcaat
     7261 ccatcactgg tctttatgaa acacgcattg atttgagtca gctaggaggt gacgctgcta
     7321 acgacgaaaa ctacgctctg gccgcctaat attacccggc atgacaggag taaaaatggc
     7381 tatcgacgaa aacaaacaga aagcgttggc ggcagcactg ggccagattg agaaacaatt
     7441 tggtaaaggc tccatcatgc gcctgggtga agaccgttcc atggatgtgg aaaccatctc
     7501 taccggttcg ctttcactgg atatcgcgct tggggcaggt ggtctgccga tgggctgcat
     7561 cgtcgaaatc tacggaccgg aatcttccgg taaaaccacg ctgacgctgc aggtgatcgc
     7621 cgcagcgcag cgtgaaggta aaacctgtgc gtttatcgat gctgaacacg cgctggaccc
     7681 aatctacgca cgtaaactgg gcgtcgatat cgacaacctg ctgtgctccc agccggacac
     7741 cggcgagcag gcactggaaa tctgtgacgc cctggcgcgt tctggcgcag tagacgttat
     7801 cgtcgttgac tccgtggcgg cactgacgcc gaaagcggaa atcgaaggcg aaatcggcga
     7861 ctct
//
