LOCUS Exported 5582 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses drosophila TAK1 tagged with V5 and 6xHis epitopes, under control of metallothionein promoter. ACCESSION . VERSION . KEYWORDS pMT-TAK1-V5-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5582) TITLE pMT-IMD-V5-His JOURNAL Unpublished REFERENCE 2 (bases 1 to 5582) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5582 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 364..386 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 378..395 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 407..833 /label=MT promoter /note="Drosophila metallothionein promoter" CDS 2974..3015 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 3025..3042 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3065..3082) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3084..3218 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3108..3127) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3162..3181 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3362..3378) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3362..3378) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3375..3397) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3386..3402 /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)." promoter complement(3410..3440) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3455..3476 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3593..3610) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3764..4352) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3844..3863) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4523..5383) /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 5146..5165 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5384..5488) /gene="bla" /label=AmpR promoter primer_bind 5556..5574 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccag tgaattaatt cgttgcagga 421 caggatgtgg tgcccgatgt gactagctct ttgctgcagg ccgtcctatc ctctggttcc 481 gataagagac ccagaactcc ggccccccac cgcccaccgc cacccccata catatgtggt 541 acgcaagtaa gagtgcctgc gcatgcccca tgtgccccac caagagtttt gcatcccata 601 caagtcccca aagtggagaa ccgaaccaat tcttcgcggg cagaacaaaa gcttctgcac 661 acgtctccac tcgaatttgg agccggccgg cgtgtgcaaa agaggtgaat cgaacgaaag 721 acccgtgtgt aaagccgcgt ttccaaaatg tataaaaccg agagcatctg gccaatgtgc 781 atcagttgtg gtcagcagca aaatcaagtg aatcatctca gtgcaactaa aggggggatc 841 tagatcgggg tacctactag tccagtgtgg tggaattcca ccatggccac agcatcgctg 901 gacgcactgc aggcagccta tgtggacttc agtgagataa cactaagaga gaaagtcggc 961 catgggtcct acggagtggt ctgcaaggcc gtttggcgcg acaagctggt tgccgtcaag 1021 gagttcttcg ccagcgccga gcagaaggac atcgagaagg aggtgaagca gttgtcgcgc 1081 gtgaagcacc cgaacatcat cgctctgcac gggatatcct cgtaccagca ggccacctac 1141 ctgataatgg agttcgccga aggtggatcg ctgcacaact tccttcacgg caaggtgaag 1201 ccggcatatt ctctggccca cgccatgagc tgggcgcgcc aatgtgcaga gggtctggca 1261 tatttgcatg ccatgacgcc aaaaccacta atacatcgcg acgtgaagcc gctgaacctg 1321 ctcttgacca acaagggacg caatctgaag atatgcgact tcggcacggt ggcggacaag 1381 tcgaccatga tgaccaacaa tcgcggcagt gccgcttgga tggcgcccga ggtcttcgaa 1441 ggctccaagt atacggagaa gtgtgacatt tttagctggg ccattgttct atgggaggtt 1501 ctgtccagga agcagccctt taaaggcatc gacaatgcct acaccatcca gtggaagatc 1561 tacaagggtg aacgcccgcc gctgctgacc acttgcccca agcgcatcga ggacctgatg 1621 accgcctgct ggaaaacggt gcccgaggat cgcccgtcga tgcagtacat agtgggcgtt 1681 atgcacgaga tcgtcaagga ctatacgggg gcggacaagg ccctggaata cacgtttgtt 1741 aatcaacaga ttgtcaccaa agagagcgac ggcacggtgg ccgctcaacc ggatagcctc 1801 agttcgcagg agggggaact gagcccctcg tccacacagt taacaccgac aacggcggcc 1861 aacgccaatg tgaacgcgat agcaatatca aaaacaacga ctagctcaat gaccgaaaat 1921 acctcatcaa catcatcgga catcacgccg acgaactcgg gccaactgga caataatccg 1981 ctattctata tggtcaccaa tcgctgggac gcgattcccg aggaggagag caacgagagc 2041 cggaacgata gcttcaacct cacctcgtcg gctgaggcca ctcagcgcct cgaaacgatc 2101 cggaacggca tgatcctgat ggcctgcaag cccatggagc agctcaccct cgacgtggag 2161 gcgaatggct ttgatctgag tcccagcgaa agcagcagca gcagcacgaa cgcaaagagc 2221 gatggccgcg aacgactcac ggtgacggac accaagccgg tgatgatgac cacggatctg 2281 tccaacaaca acggcggcat ccacgcccac tcgaacggac tgctgagcca tgcgaatggt 2341 tggcaagcaa gagatgagga gctgcaggag caagagcatg agcaggagat tgtcaactcg 2401 ttggacgtcg acgtggatcc cgacgaggat gagaacgacg gcaccgaaca gtcactggcc 2461 gagattcttg atccggagct ccagccagag ccgccgatac ccaacgatgc cgaatcgcag 2521 ctcatctacc gggaccaccg acacatggcc aaggagtacc tgagcgtcga cacgaacctc 2581 tactacgcgc aggacttcaa ggacaagctc atcgtgcaga tggaccgaac cgagcgcgaa 2641 cagaagcagg agcttctgcg caagatgaag gacaaggagg gtcttcagag tctttacaac 2701 aatctgcagc agcagtacgc ttcacgccaa cttgcggccg gccatcatcc gcaacctcat 2761 cctcatccgc atccaaatca gcttcagcat ccacactcac atccgcctat gcatttcctg 2821 caggatgagg gctgtggact gctgcccgga tcggtgtgcg gaggctctga gtccgtggaa 2881 gaaggctggg tggtcatccc accgcatcac aatgcggaat tctgcagata tccagcacag 2941 tggcggccgc tcgagtctag agggcccttc gaaggtaagc ctatccctaa ccctctcctc 3001 ggtctcgatt ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga 3061 tcagcctcga ctgtgccttc taagatccag acatgataag atacattgat gagtttggac 3121 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg 3181 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt 3241 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca 3301 aatgtggtat ggctgattat gatcagtcga cctgcaggca tgcaagcttg gcgtaatcat 3361 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 3421 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 3481 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 3541 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 3601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 3661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 3721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 3781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 3841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 3901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 3961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 4021 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 4081 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 4141 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 4201 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 4261 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 4321 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 4381 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 4441 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 4501 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 4561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 4621 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 4681 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 4741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 4801 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 4861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 4921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 4981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 5041 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 5101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 5161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 5221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 5281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 5341 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 5401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 5461 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct 5521 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc 5581 gt //