LOCUS Exported 10925 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION parental, all-in-one CRISPRi vector for B. burgdorferi, parental for gRNA cloning. ACCESSION . VERSION . KEYWORDS pBbdCas9B(RBSmut)_arr2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10925) AUTHORS Takacs CN, Scott M, Chang Y, Kloos ZA, Irnov I, Rosa PA, Liu J, Jacobs-Wagner C TITLE A CRISPR interference platform for selective downregulation of gene expression in Borrelia burgdorferi. JOURNAL Appl Environ Microbiol. 2020 Nov 30. pii: AEM.02519-20. doi: 10.1128/AEM.02519-20. PUBMED 33257311 REFERENCE 2 (bases 1 to 10925) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10925 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 2346..2390 /label=T5 promoter /note="bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator" protein_bind 2366..2382 /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)." protein_bind 2398..2414 /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)." CDS 2451..6557 /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 TILERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" rep_origin 7251..7839 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7740..7759 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 aaagccctaa gtcttttaat aaaaaatttt ttgtaaaaaa gttggcaaaa atagtttttg 61 ctatatattt atatataaga aaaaattcaa tactcaagga gataaaaaat ggaaaactta 121 aacaataatc cacaaaaaat taatcaaata aataagcaaa ctaattttat tggttttgaa 181 aattttatag aacttgataa tcaagttaat caaactaaag aaaaaagtaa attaagtaag 241 ataggtaaga aattacccgg aataagcagt caagtctagt gcggccgcat ggcttgttat 301 gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc 361 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct 421 aaaacaaagt taaacatcat gaggaggtaa cgtcatggta aaagattgga ttcctatttc 481 tcatgataat tataaacaag tacaaggacc attttatcat ggaactaaag ctaatttagc 541 aattggtgat ttactaacta caggatttat ttctcatttt gaagatggtc gaattcttaa 601 acatatttat ttttcagctt taatggaacc agcagtttgg ggagctgaac ttgctatgtc 661 actatctggt cttgaaggtc gtggttatat atatatagtt gaaccaacag gaccatttga 721 agatgatcca aatcttacaa ataaaaaatt tcctggtaat ccaacacaat cttatagaac 781 ttgtgaacct ttaagaattg ttggtgttgt tgaagattgg gaaggacatc ctgttgaatt 841 aataagagga atgttagatt cattagaaga tttaaaacgt cgtggtttac atgttattga 901 agattaacct ggcgcgccaa ataatgcccg tgtccctgtt gaatagggac acggtttttt 961 aacaattttt ttattcctag gccttagtgg ccggtacccg gggatcctta ttactggccg 1021 ctttctagcc tggatacttg tctggctagt tgcattagag agtcagctag agccctaggg 1081 ctagctgttt gtgtattcgg agccagtgta gtttttcttt ttactaacga tacgggtaat 1141 aattgattgc cttttacagc ctgaccttga gacaattgca aaagtctgtc cacagaggtt 1201 tgacctaata atctaaaatc ttgtttgata gtagtaagtg gaggtatgta gcaactagaa 1261 tcttctgtat catcataacc cacaactgaa atatcagccc caactctaag tccactttca 1321 gtaatagctc gcatagctcc taaagccatt tgatcattag caactaacat agcagtaggg 1381 acaatgcctt catttaacat ttgcattgtt tgctgaaatc ctgacatagc agaccaatct 1441 ccttctcttt ctgctattgg ttgtatttga tttcttgtta aatacttgtg ccatcctgct 1501 aatctcaatc tggcacttac tgaacttaag ggacctgcaa gtagtgcaat ttgttgatgg 1561 cctaatgcaa caaggtgttc aactcctaac cttgttccat cttcgtgaga aaaaattata 1621 gagtttattg gggtttggtc cgacacatcc aaaaacaatg ctggaacatt agtacatgct 1681 gcttcaactg caatcgcatc ttggtcatca agaggataat taattataag tccactaaca 1741 cgttgagcta ataaattatg tactgctgcc ttacaagcct ctacacctga tctctcaacc 1801 attgatacaa caacgcttgc accaagttga tcagctctcg atttaattgc ggctacaatt 1861 tgactaggcg catgtaatgc aagtgatgat gtggcaactc caattaaaag tgattgtttc 1921 cccgcaagtt gctgagcaac gcgatttggt atataattca actctgccat agctgcttca 1981 accttctctc ttgtttttgc agatacatga cttgcttggt tcacaactct cgaaaccgtt 2041 tggtagctaa ccccggcata ttcagcaaca tcatacaaag taacaggttt catggatcat 2101 tcctccatga taaaatttaa atttctgact ttggcaaatc cttttgcctg ttaaaaagaa 2161 tagcacaaaa attaaaaaaa aagaaacgca aatagcaata gcattaaaaa taattattat 2221 tatttatttc tttgattaaa atcaaaagcc tttaatgtat tttcttgatc tttcatgcgc 2281 ttaacggtaa aatccaaggt attaagttca ccttccactt ttctctctct atcttctcta 2341 gaaaatcata aaaaatttat ttgctttgtg agcggataac aattataata gattcaattg 2401 tgagcggata acaatttcac acagaattca ttaaagaaga gaaattacat atggataaga 2461 aatactcaat aggcttagct atcggcacaa atagcgtcgg atgggcggtg atcactgatg 2521 aatataaggt tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca 2581 aaaaaaatct tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgtc 2641 tcaaacggac agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg 2701 agattttttc aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt 2761 cttttttggt ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag 2821 atgaagttgc ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag 2881 attctactga taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt 2941 ttcgtggtca ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac 3001 tatttatcca gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa 3061 gtggagtaga tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa 3121 atctcattgc tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt 3181 tgtcattggg tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat 3241 tacagctttc aaaagatact tacgatgatg atttagataa tttattggcg caaattggag 3301 atcaatatgc tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag 3361 atatcctaag agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac 3421 gctacgatga acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc 3481 cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg 3541 atgggggagc tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg 3601 atggtactga ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga 3661 cctttgacaa cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga 3721 gaagacaaga agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct 3781 tgacttttcg aattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat 3841 ggatgactcg gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata 3901 aaggtgcttc agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa 3961 atgaaaaagt actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat 4021 tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac 4081 agaagaaagc cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat 4141 taaaagaaga ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg 4201 aagatagatt taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata 4261 aagatttttt ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga 4321 ccttatttga agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg 4381 atgataaggt gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc 4441 gaaaattgat taatggtatt agggataagc aatctggcaa aacaatatta gattttttga 4501 aatcagatgg ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat 4561 ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata 4621 ttgcaaattt agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg 4681 ttgatgaatt ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg 4741 cacgtgaaaa tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa 4801 tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata 4861 ctcaattgca aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg 4921 tggaccaaga attagatatt aatcgtttaa gtgattatga tgtcgatgcc attgttccac 4981 aaagtttcct taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc 5041 gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga 5101 gacaacttct aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg 5161 aacgtggagg tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa 5221 ctcgccaaat cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg 5281 atgaaaatga taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt 5341 ctgacttccg aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg 5401 cccatgatgc gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac 5461 ttgaatcgga gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta 5521 agtctgagca agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga 5581 acttcttcaa aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg 5641 aaactaatgg ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc 5701 gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa aacagaagtc cagacaggcg 5761 gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa 5821 aagactggga tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc 5881 tagtggttgc taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac 5941 tagggatcac aattctggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag 6001 ctaaaggata taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt 6061 ttgagttaga aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa 6121 atgagctggc tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa 6181 agttgaaggg tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc 6241 attatttaga tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag 6301 atgccaattt agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg 6361 aacaagcaga aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt 6421 ttaaatattt tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag 6481 atgccactct tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc 6541 agctaggagg tgactaataa aagcttgatc agatcccagc ttttttttga agtgcctggc 6601 agtaagttga aaaaatttga aataaacttg gactatgtta ataaaaaaat acaaattaac 6661 gaacaaaaag atttgaatta ttatataatt tagttaaaag caattttaaa tgagggaggt 6721 ttccatcata tggctaaacc tttaagtcaa gaagaaagta ctttaattga aagagctact 6781 gctactatta atagtattcc tattagtgaa gattatagtg ttgctagtgc tgctttaagt 6841 agtgatggta gaatttttac tggtgttaat gtttatcatt ttactggtgg tccttgtgct 6901 gaattagttg ttttaggtac tgccgccgct gctgctgctg gtaatctcac ttgtattgtt 6961 gctattggta atgaaaatag aggtatttta agtccttgtg gtagatgtag acaagtttta 7021 ttagatttac atcctggtat taaagctatt gttaaagata gtgatggtca acctactgct 7081 gttggtatta gagaattatt acctagtggt tatgtttggg aaggttaatg acgtcgccgg 7141 agcggtcgag ttctggaccg accggctcgg gttctcatga ccaaaatccc ttaacgtgag 7201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 7261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 7321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 7381 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 7441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 7501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 7561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 7621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 7681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 7741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 7801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 7861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc actagatccc 7921 cggcaagttc atccttaggg ttttcatatc tcttaaaaag tcaatttctg cttgattaat 7981 ttcttgtgga ttattgtttt tgcggttttt cattttttat ctccttgagt attgaatttt 8041 ttcttatata taaatatata gcaaaaacta tttttgccaa cttttttaca aaaaattttt 8101 tattaaaaca cttagggctt tgctaaattc tcttttaaaa gaacttagta aagccctatg 8161 gatttaagaa ctgattatac ttacgtagta aaaatactac agatattaat agtatagacc 8221 atatcaaaat cataatcaat ttatatttat ctaaaattgt agtccataca aaatacaaaa 8281 aaattaggtt tttctaaatt ctattttaaa agaacttagt aacacctaat ctctaaatta 8341 cttacaaaat tctaaaatta gatttttagt tcttcatatt tctttaaaag ttttttaaga 8401 aaatcttttt gattttcaaa aatttcatgc atcataaaac tagtaaattt ggattttttt 8461 ttgtaaaaat cataactttc ttgggtttta agttgaaatc ttaatggttt tattggattt 8521 tgcctagatt tttttaacgc cggactttct ttatctttta taaaattata agaattttta 8581 ataccatttt ctattaaata atcttctttt ataagcccct cttgcagtgc agttgctaat 8641 cttaaatact tatacgcttg agtttttgct acatcaaagt cttttataaa atcaagaaaa 8701 gttttataac catcaaattt ataatattga ttatctttta tttcttttaa aattttaata 8761 ttttccaccc tgtggaatat ttcttttttg aaattgtcat ttaacttttc ctttagtgtc 8821 ttgtaataag aaattttact ttcattaatc attaaatttt gttcaacata agtatctatc 8881 ttttttgtta aaactttaac aacacgtttt ttttccataa aaattttcct ttatttttgc 8941 aacattttcc ttaattgata taattgttac tattttagat tttttaatat ccacttaaga 9001 attaatccct cactagaata ttttttatta tagaattcat atacatgttt ttctaaatct 9061 ataaatttat caacaattgc tttattatag ggtgtatcta tcttttcttt tcttaataat 9121 cgtctcatat tctttaaata acaacaaaca ctgccggttt taaacctaaa ttctatgtaa 9181 tatatttttg aaaaagtata tacattttca accccgttta tttcatactt tacaaaaaca 9241 ttatttattg gttttctata accatagtaa atgcctaaaa atttgtcccc gtctttcata 9301 ggatatagat aaagtttttg aatctctgat ttatctaata atttttcaaa aaaaattata 9361 caccgctttt gtttgttact aactactcta tgcaatgcct ttaatatttt tgtgtgatat 9421 atatttttat cacctttttt ttctttcagc aggaagatac tttcttttgg aaatttaatt 9481 ttttgttcat ttattcttgc agccttaagt ccgattttca tatttaatcc ttatatagcc 9541 tttctattta attcttttaa actcaataaa cttttttggt ttttcattag ttctaataat 9601 tcgcaataat atgtgcaaaa cactttattg tattctattt tcttttggtt attcaaatat 9661 tcttttataa ttggctttag aagttcaata tttacttctt tttttagttg ttcaataagg 9721 atattgaaaa tgtttgtttt taaatcttga taattttgtt gagaattttc tttttttctt 9781 tcaattgact tctctaattt acgctttata ttgtttaaat cgctatattt atgactttca 9841 ataataaaat gaggcttaaa tttgtaattt tcgtatattt tttggaaact tgtttctaat 9901 tgtttagaat tatattcatt tttttcaaat tctttttgag tgttgcatag aatttccttt 9961 aatttttctt gtttttcttt catacaagat ttagtaaaat gtatattttt attttttagt 10021 aatttaattt ctattatttt tatgatttta agtattttga ttatattatc tttatcaata 10081 tttaaattga ataaaattgt aagaaaaatt gaaagaattt ttttacataa aaagttacat 10141 ttattgaagt aatttcttac ttgatacttt tctatttcat taatctttct ttcttctttt 10201 atattatttt tattatttaa acactccttt aaatctacac tcccattttt agtaaatcta 10261 tctttaaggc cccttgaaac tctattttga aatcttaaat cttttttttc ttgaaagtat 10321 ttatagattt tttggtaaca ttcttttttt gaaaaattaa gttgatagta tatttcggtc 10381 cccaaattta cccctaagtg tttgtgataa ttagttgtga ctttaaattc tttttctaat 10441 ttgtaaaggt aattttgcag tgtttttagt ttaacgggtg tttgactatt tcttttaagg 10501 ttttcattaa agtaatagag tatgtttttt tgtgtgtatt tcttatatgt tttatttatg 10561 tacgctagtg ttgagataag gactactaat ttatactgta atttattgta gcaagttgtg 10621 gttttactgg ttttgtttat attagataag ttttccattt atagctcctt gaataccgga 10681 ttttttatct ataaagtata tatatcacaa ttttgattta aaagtaaatt ctttttttaa 10741 aaatacaaaa attttttata aattatatag accatgatat tgtattaata ctaattgatt 10801 ttaatttaag gttgaactaa aataaacata gttttgtaga aaataatttt tcattatttt 10861 gctacttgat attggatcgt aaaaatatta gggctttaca aagttctttt aaaagagaat 10921 ttagt //