LOCUS Exported 5064 bp ds-DNA circular SYN 04-JAN-2022 DEFINITION Tetrazine 3.0 tRNA synthetase and cognate amber suppressing tRNA derived from M. barkeri for expression in E.coli.. ACCESSION . VERSION . KEYWORDS pDule-Tet3.0 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5064) AUTHORS Jang HS, Jana S, Blizzard RJ, Meeuwsen JC, Mehl RA TITLE Access to Faster Eukaryotic Cell Labeling with Encoded Tetrazine Amino Acids. JOURNAL J Am Chem Soc. 2020 Apr 22;142(16):7245-7249. doi: 10.1021/jacs.9b11520. Epub 2020 Apr 10. PUBMED 32251579 REFERENCE 2 (bases 1 to 5064) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 4, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5064 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1513..1542 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1558..1575 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1631..1648) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1801..1887 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind complement(2094..2111) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2228..2773) /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(2308..2327) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2885..2913 /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" CDS 2961..4151 /codon_start=1 /gene="tet" /product="tetracycline efflux protein" /label=TcR /note="confers resistance to tetracycline" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" primer_bind complement(3313..3332) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 4458..4480 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4472..4489 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4473..4489 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 gccgttatac gttgtttacg ctttgaggaa tcccatatga tggataaaaa accgctggat 61 gtgctgatta gcgcgaccgg cctgtggatg agccgtaccg gcaccctgca taaaatcaaa 121 catcatgaag tgagccgcag caaaatctat attgaaatgg cgtgcggcga tcatctggtg 181 gtgaacaaca gccgtagctg ccgtaccgcg cgtgcgtttc gtcatcataa ataccgcaaa 241 acctgcaaac gttgccgtgt gagcgatgaa gatatcaaca actttctgac ccgtagcacc 301 gaaagcaaaa acagcgtgaa agtgcgtgtg gtgagcgcgc cgaaagtgaa aaaagcgatg 361 ccgaaaagcg tgagccgtgc gccgaaaccg ctggaaaata gcgtgagcgc gaaagcgagc 421 accaacacca gccgtagcgt tccgagcccg gcgaaaagca ccccgaacag cagcgttccg 481 gcgtctgcgc cggcaccgag cctgacccgc agccagctgg atcgtgtgga agcgctgctg 541 tctccggaag ataaaattag cctgaacatg gcgaaaccgt ttcgtgaact ggaaccggaa 601 ctggtgaccc gtcgtaaaaa cgattttcag cgcctgtata ccaacgatcg tgaagattat 661 ctgggcaaac tggaacgtga tatcaccaaa ttttttgtgg atcgcggctt tctggaaatt 721 aaaagcccga ttctgattcc ggcggaatat gtggaacgta tgggcattaa caacgacacc 781 gaactgagca aacaaatttt ccgcgtggat aaaaacctgt gcctgcgtcc gatgctggcc 841 ccgaccggtt ataactatct gcgtaaactg gatcgtattc tgccgggtcc gatcaaaatt 901 tttgaagtgg gcccgtgcta tcgcaaagaa agcgatggca aagaacacct ggaagaattc 961 accatggttg gttttgctca aatgggcagc ggctgcaccc gtgaaaacct ggaagcgctg 1021 atcaaagaat tcctggatta tctggaaatc gacttcgaaa ttgtgggcga tagctgcatg 1081 gtgtatggcg ataccctgga tattatgcat ggcgatctgg aactgagcag cgcggtggtg 1141 ggtccggtta gcctggatcg tgaatggggc attgataaac cgtggattgg cgcgggtttt 1201 ggcctggaac gtctgctgaa agtgatgcat ggcttcaaaa acattaaacg tgcgagccgt 1261 agcgaaagct actataacgg cattagcacg aacctgtaac tgcagtttca aacgctaaat 1321 tgcctgatgc gctacgctta tcaggctaca tgatctctgc aatatattga gtttgcgtgc 1381 ttttgtaggc cggataaggc gttcacgccg catccggcag gaaacagcaa acaatccaaa 1441 acgccgcgtt cagcggcgtt ttttctgcgt ttcttcgcga attaattccg ctctcgagcg 1501 aagcttgggc ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat 1561 catcatcatc atcattgagt ttaaacggtc tccagcttgg ctgttttggc ggatgagaga 1621 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt 1681 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac 1741 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat 1801 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg 1861 gtgaacgctc tcctgagtag gacaaatccg ccgggagctg tccctcctgt tcagctactg 1921 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact 1981 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa 2041 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc 2101 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc 2161 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa 2221 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc 2281 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc 2341 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc 2401 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac 2461 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt 2521 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt 2581 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg 2641 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt 2701 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagttta cgcgcagacc 2761 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca 2821 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc 2881 atgtttgaca gcttatcatc gataagcttt aatgcggtag tttatcacag ttaaattgct 2941 aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg 3001 tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg 3061 atatcgtcca ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt 3121 tgatgcaatt tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc 3181 cagtcctgct cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac 3241 ccgtcctgtg gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg 3301 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg 3361 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt 3421 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc 3481 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga ccgatgccct 3541 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg 3601 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg 3661 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg 3721 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt 3781 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc 3841 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg 3901 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg 3961 gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcatt ggaccgctga 4021 tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag 4081 gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca 4141 cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg 4201 agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc 4261 catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggctcct tgcatgcacc 4321 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca 4381 ggagtcgcat aagggagagc gtctggcgaa agggggatgt gctgcaaggc gattaagttg 4441 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagcttaa 4501 aaaaaatcct tagctttcgc taagatctgc agtggcggaa accccgggaa tctaacccgg 4561 ctgaacggat ttagagtcca ttcgatctac atgatcaggt tcccgcggcc gcgaattcag 4621 cgttacaagt attacacaaa gttttttatg ttgagaatat ttttttgatg gggcgccact 4681 tatttttgat cgttcgctca aagaagcggc gccagggttg tttttctttt caccagtgag 4741 acgggcaaca gaacgccatg agcggcctca tttcttattc tgagttacaa cagtccgcac 4801 cgctgccggt agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac 4861 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgcccaaca gtcccccggc 4921 cacggggcct gccaccatac ccacgccgaa acaagcgccc tgcaccatta tgttccggat 4981 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acggtacctc gggttgtcag 5041 cctgtcccgc ttataagatc atac //