LOCUS       sequence-459808-        8535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aTc inducible knockdown of Fis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8535)
  AUTHORS   Deyell M, Opuu V, Griffiths AD, Tans SJ, Nghe P
  TITLE     Global regulators enable bacterial adaptation to a phenotypic 
            trade-off.
  JOURNAL   iScience. 2024 Dec 9;28(1):111521. doi: 10.1016/j.isci.2024.111521. 
            eCollection 2025 Jan 17.
  PUBMED    39811663
REFERENCE   2  (bases 1 to 8535)
  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: "iScience.";
            date: "2024-12-9"; volume: "28(1):111521. doi"; pages: " 
            10.1016/j.isci.2024.111521. eCollection 2025 Jan 17"
COMMENT     Sequence Label: pKDcas9 fis cerulean
FEATURES             Location/Qualifiers
     source          1..8535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..546
                     /direction=RIGHT
                     /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     447..466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     663..680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(708..735)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(751..822)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(846..2432,2433..2435,2436..4922,4923..4925,
                     4926..4952))
                     /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
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     RBS             4959..4970
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    4989..5007
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        5015..5069
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    5015..5033
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             5085..5711
                     /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     complement(5161..5180)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    5727..5858
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   5859..5894
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   5925..5960
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_RNA        6081..6159
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     primer_bind     complement(6304..6322)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             6501..7295
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6555..6574)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7165..7184
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      7314..7408
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(7501..7528)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7577..7606)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(8017..8028)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             8346..8357
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(8361..8395)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
ORIGIN
        1 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
       61 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
      121 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
      181 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
      241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
      301 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
      361 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
      421 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
      481 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
      541 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
      601 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
      661 gtagcgagtc agtgagcgag gaagcggaat atatccctag gtataaacgc agaaaggccc
      721 acccgaaggt gagccagtgt gactctagta gagagcgttc accgacaaac aacagataaa
      781 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg agatccttac
      841 tcgagttagt cacctcctag ctgactcaaa tcaatgcgtg tttcataaag accagtgatg
      901 gattgatgga taagagtggc atctaaaact tcttttgtag acgtatatcg tttacgatca
      961 attgttgtat caaaatattt aaaagcagcg ggagctccaa gattcgtcaa cgtaaataaa
     1021 tgaataatat tttctgcttg ttcacgtatt ggtttgtctc tatgtttgtt atatgcacta
     1081 agaactttat ctaaattggc atctgctaaa ataacacgct tagaaaattc actgatttgc
     1141 tcaataatct catctaaata atgcttatgc tgctccacaa acaattgttt ttgttcgtta
     1201 tcttctggac tacccttcaa cttttcataa tgactagcta aatataaaaa attcacatat
     1261 ttgcttggca gagccagctc atttcctttt tgtaattctc cggcactagc cagcatccgt
     1321 ttacgaccgt tttctaactc aaaaagacta tatttaggta gtttaatgat taagtctttt
     1381 ttaacttcct tatatccttt agcttctaaa aagtcaatcg gatttttttc aaaggaactt
     1441 ctttccataa ttgtgatccc tagtaactct ttaacggatt ttaacttctt cgatttccct
     1501 ttttccacct tagcaaccac taggactgaa taagctaccg ttggactatc aaaaccacca
     1561 tatttttttg gatcccagtc ttttttacga gcaataagct tgtccgaatt tctttttggt
     1621 aaaattgact ccttggagaa tccgcctgtc tgtacttctg ttttcttgac aatattgact
     1681 tggggcatgg acaatacttt gcgcactgtg gcaaaatctc gccctttatc ccagacaatt
     1741 tctccagttt ccccattagt ttcgattaga gggcgtttgc gaatctctcc atttgcaagt
     1801 gtaatttctg ttttgaagaa gttcatgata ttagagtaaa agaaatattt tgcggttgct
     1861 ttgcctattt cttgctcaga cttagcaatc attttacgaa catcataaac tttataatca
     1921 ccatagacaa actccgattc aagttttgga tatttcttaa tcaaagcagt tccaacgacg
     1981 gcatttagat acgcatcatg ggcatgatgg taattgttaa tctcacgtac tttatagaat
     2041 tggaaatctt ttcggaagtc agaaactaat ttagatttta aggtaatcac tttaacctct
     2101 cgaataagtt tatcattttc atcgtattta gtattcatgc gactatccaa aatttgtgcc
     2161 acatgcttag tgatttggcg agtttcaacc aattggcgtt tgataaaacc agctttatca
     2221 agttcactca aacctccacg ttcagctttc gttaaattat caaacttacg ttgagtgatt
     2281 aacttggcgt ttagaagttg tctccaatag tttttcatct ttttgactac ttcttcactt
     2341 ggaacgttat ccgatttacc acgattttta tcagaacgcg ttaagacctt attgtctatt
     2401 gaatcgtctt taaggaaact ttgtggaaca atggcatcga catcataatc acttaaacga
     2461 ttaatatcta attcttggtc cacatacatg tctcttccat tttggagata atagagatag
     2521 agcttttcat tttgcaattg agtattttca acaggatgct ctttaagaat ctgacttcct
     2581 aattctttga taccttcttc gattcgtttc atacgctctc gcgaattttt ctggcccttt
     2641 tgagttgtct gattttcacg tgccatttca ataacgatat tttctggctt atgccgcccc
     2701 attactttga ccaattcatc aacaactttt acagtctgta aaataccttt tttaatagca
     2761 gggctaccag ctaaatttgc aatatgttca tgtaaactat cgccttgtcc agacacttgt
     2821 gctttttgaa tgtcttcttt aaatgtcaaa ctatcatcat ggatcagctg cataaaattg
     2881 cgattggcaa aaccatctga tttcaaaaaa tctaatattg ttttgccaga ttgcttatcc
     2941 ctaataccat taatcaattt tcgagacaaa cgtccccaac cagtataacg gcgacgttta
     3001 agctgtttca tcaccttatc atcaaagagg tgagcatatg ttttaagtct ttcctcaatc
     3061 atctccctat cttcaaataa ggtcaatgtt aaaacaatat cctctaagat atcttcattt
     3121 tcttcattat ccaaaaaatc tttatcttta ataattttta gcaaatcatg gtaggtacct
     3181 aatgaagcat taaatctatc ttcaactcct gaaatttcaa cactatcaaa acattctatt
     3241 tttttgaaat aatcttcttt taattgctta acggttactt ttcgatttgt tttgaagagt
     3301 aaatcaacaa tggctttctt ctgttcacct gaaagaaatg ctggttttcg cattccttca
     3361 gtaacatatt tgacctttgt caattcgtta taaaccgtaa aatactcata aagcaaacta
     3421 tgttttggta gtactttttc atttggaaga tttttatcaa agtttgtcat gcgttcaata
     3481 aatgattgag ctgaagcacc tttatcgaca acttcttcaa aattccatgg ggtaattgtt
     3541 tcttcagact tccgagtcat ccatgcaaaa cgactattgc cacgcgccaa tggaccaaca
     3601 taataaggaa ttcgaaaagt caagattttt tcaatcttct cacgattgtc ttttaaaaat
     3661 ggataaaagt cttcttgtct tctcaaaata gcatgcagct cacccaagtg aatttgatgg
     3721 ggaatagagc cgttgtcaaa ggtccgttgc ttgcgcagca aatcttcacg atttagtttc
     3781 accaataatt cctcagtacc atccattttt tctaaaattg gtttgataaa tttataaaat
     3841 tcttcttggc tagctccccc atcaatataa cctgcatatc cgttttttga ttgatcaaaa
     3901 aagatttctt tatacttttc tggaagttgt tgtcgaacta aagcttttaa aagagtcaag
     3961 tcttgatgat gttcatcgta gcgtttaatc attgaagctg ataggggagc cttagttatt
     4021 tcagtattta ctcttaggat atctgaaagt aaaatagcat ctgataaatt cttagctgcc
     4081 aaaaacaaat cagcatattg atctccaatt tgcgccaata aattatctaa atcatcatcg
     4141 taagtatctt ttgaaagctg taatttagca tcttctgcca aatcaaaatt tgatttaaaa
     4201 ttaggggtca aacccaatga caaagcaatg agattcccaa ataagccatt tttcttctca
     4261 ccggggagct gagcaatgag attttctaat cgtcttgatt tactcaatcg tgcagaaaga
     4321 atcgctttag catctactcc acttgcgtta atagggtttt cttcaaataa ttgattgtag
     4381 gtttgtacca actggataaa tagtttgtcc acatcactat tatcaggatt taaatctccc
     4441 tcaatcaaaa aatgaccacg aaacttaatc atatgcgcta aggccaaata gattaagcgc
     4501 aaatccgctt tatcagtaga atctaccaat ttttttcgca gatgatagat agttggatat
     4561 ttctcatgat aagcaacttc atctactata tttccaaaaa taggatgacg ttcatgcttc
     4621 ttgtcttctt ccaccaaaaa agactcttca agtcgatgaa agaaactatc atctactttc
     4681 gccatctcat ttgaaaaaat ctcctgtaga taacaaatac gattcttccg acgtgtatac
     4741 cttctacgag ctgtccgttt gagacgagtc gcttccgctg tctctccact gtcaaataaa
     4801 agagccccta taagattttt tttgatactg tggcggtctg tatttcccag aaccttgaac
     4861 tttttagacg gaaccttata ttcatcagtg atcaccgccc atccgacgct atttgtgccg
     4921 atagctaagc ctattgagta tttcttatcc atagatcctt tctcctcttt agatcttttg
     4981 aattcttttc tctatcactg atagggaggg tcaatctcta tcactgatag ggagtggtaa
     5041 aataactcta tcaacgatag agtgtcaaca aaaattagga attaatgatg tctagattag
     5101 ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt
     5161 taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg
     5221 taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata
     5281 ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa
     5341 gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc
     5401 ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt
     5461 tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact ttaggttgcg
     5521 tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata
     5581 gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt gcagagccag
     5641 ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa
     5701 gtgggtctta aggtaccatg ggatcctatt tcttaataac taaaaatatg gtataatact
     5761 cttaataaat gcagtaatac aggggctttt caagactgaa gtctagctga gacaaatagt
     5821 gcgattacga aattttttag acaaaaatag tctacgaggt tttagagcta tgctgttttg
     5881 aatggtccca aaacgtatat tacgcatttt ttgcacgaga tgaagtttta gagctatgct
     5941 gttttgaatg gtcccaaaac ttcagcacac tgagacttgt tgagttgaat tcatgagtat
     6001 taagtattgt tttatggctg ataaatttct ttgaatttct ccttgattat ttgttataaa
     6061 agttataaaa taatcttgtt ggaaccattc aaaacagcat agcaagttaa aataaggcta
     6121 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttgata cttctattct
     6181 actctgactg caaaccaaaa aaacaagcgc tttcaaaacg cttgttttat catttttagg
     6241 gaaattaatc tcttaatcct tttatcattc tacatttagg cgctgccatc ttaagacgaa
     6301 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6361 cgtcggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     6421 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga
     6481 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     6541 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     6601 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     6661 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     6721 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     6781 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6841 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6901 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6961 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     7021 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     7081 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     7141 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     7201 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     7261 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctctcgc ttggactcct
     7321 gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg
     7381 ccgccgggcg ttttttattg gtgagaatcc aagcactagt tggtaccggc atcaatagta
     7441 aaataataaa aaagccggat taataatctg gctttttata ttctctctct agtatataaa
     7501 cgcagaaagg ccaacccgaa ggtgagccag tgtgagacta tcatactcat atcgtgtgaa
     7561 gatttgctgc gctattcaga tcctcttctg agatgagttt ttgttcgggc ccaagcttct
     7621 cacttataca attcatccat gccgagggta ataccggccg ctgtcacaaa ctccaggaga
     7681 accatatggt cacgtttttc gtttgggtct ttagagagtt ttgactgcgt cgacaggtaa
     7741 tggttgtcag ggagcagtac cggcccatcg ccgatcgggg tattctgctg gtagtgatcc
     7801 gccagttgca cgctcccatc ttcgatattg tggcgaattt taaagtttgc cttgatgcca
     7861 tttttctgtt tatctgcagt gatataaaca ttatcgctaa tagcgttgta ttccagcttg
     7921 tgcccaagaa tgttgccgtc ttctttgaaa tcaattcctt tcagttcgat gcgatttacc
     7981 agcgtatcgc cctcaaattt cacttccgca cgggttttgt agttcccatc gtccttaaag
     8041 aaaattgtgc gttcctgaac ataaccttcc ggcattgcag atttaaagaa gtcgtgttgt
     8101 ttcatatgat ctgggtaacg cgcaaaacac tgtacccccc acgttaatgt cgtgacaagg
     8161 gtaggccagg gcaccggcag tttgccagtc gtacagatga atttgagcgt gagcttgccg
     8221 tacgtcgcgt caccctcgcc ctcgccgctg accgaaaatt tatggccatt cacatcacca
     8281 tcaagttcaa caagaatcgg caccacgccg gtaaataatt cttcgccctt gcttaccatt
     8341 tagtatttct cctctttagt actagtatta tacctaggac tgagctagct gtcaaagatc
     8401 tttagaattc cagaaatcat ccttagcgac tagtaacaac ttatatcgta tggggctgac
     8461 ttcaggtgct acatttgaag agataaattg cactgaaatc tagaaatatt ttatctgatt
     8521 aataagatga tcttc
//
