LOCUS       sequence-459909-        8600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aTc inducible knockdown of FliZ and HNS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8600)
  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 8600)
  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 73 cerulean
FEATURES             Location/Qualifiers
     source          1..8600
                     /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..5959
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   5990..6025
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_RNA        6146..6224
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     primer_bind     complement(6369..6387)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             6566..7360
                     /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(6620..6639)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7230..7249
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      7379..7473
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(7566..7593)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7642..7671)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(8082..8093)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     RBS             8411..8422
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(8426..8460)
                     /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 aaaccgataa acagccctgc gttatatgag ttatgtttta gagctatgct
     5941 gttttaatgg tcccaaaacc attgtagtaa tctcaaactt atattggggg ttttagagct
     6001 atgctgtttt gaatggtccc aaaacttcag cacactgaga cttgttgagt tgaattcatg
     6061 agtattaagt attgttttat ggctgataaa tttctttgaa tttctccttg attatttgtt
     6121 ataaaagtta taaaataatc ttgttggaac cattcaaaac agcatagcaa gttaaaataa
     6181 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgatacttct
     6241 attctactct gactgcaaac caaaaaaaca agcgctttca aaacgcttgt tttatcattt
     6301 ttagggaaat taatctctta atccttttat cattctacat ttaggcgctg ccatcttaag
     6361 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6421 ttagacgtcg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt
     6481 aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gatctgatca
     6541 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     6601 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     6661 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6721 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     6781 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6841 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     6901 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6961 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     7021 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     7081 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     7141 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     7201 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     7261 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     7321 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ctcgcttgga
     7381 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     7441 ggttgccgcc gggcgttttt tattggtgag aatccaagca ctagttggta ccggcatcaa
     7501 tagtaaaata ataaaaaagc cggattaata atctggcttt ttatattctc tctctagtat
     7561 ataaacgcag aaaggccaac ccgaaggtga gccagtgtga gactatcata ctcatatcgt
     7621 gtgaagattt gctgcgctat tcagatcctc ttctgagatg agtttttgtt cgggcccaag
     7681 cttctcactt atacaattca tccatgccga gggtaatacc ggccgctgtc acaaactcca
     7741 ggagaaccat atggtcacgt ttttcgtttg ggtctttaga gagttttgac tgcgtcgaca
     7801 ggtaatggtt gtcagggagc agtaccggcc catcgccgat cggggtattc tgctggtagt
     7861 gatccgccag ttgcacgctc ccatcttcga tattgtggcg aattttaaag tttgccttga
     7921 tgccattttt ctgtttatct gcagtgatat aaacattatc gctaatagcg ttgtattcca
     7981 gcttgtgccc aagaatgttg ccgtcttctt tgaaatcaat tcctttcagt tcgatgcgat
     8041 ttaccagcgt atcgccctca aatttcactt ccgcacgggt tttgtagttc ccatcgtcct
     8101 taaagaaaat tgtgcgttcc tgaacataac cttccggcat tgcagattta aagaagtcgt
     8161 gttgtttcat atgatctggg taacgcgcaa aacactgtac cccccacgtt aatgtcgtga
     8221 caagggtagg ccagggcacc ggcagtttgc cagtcgtaca gatgaatttg agcgtgagct
     8281 tgccgtacgt cgcgtcaccc tcgccctcgc cgctgaccga aaatttatgg ccattcacat
     8341 caccatcaag ttcaacaaga atcggcacca cgccggtaaa taattcttcg cccttgctta
     8401 ccatttagta tttctcctct ttagtactag tattatacct aggactgagc tagctgtcaa
     8461 agatctttag aattccagaa atcatcctta gcgactagta acaacttata tcgtatgggg
     8521 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     8581 tgattaataa gatgatcttc
//
