LOCUS Exported 3839 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression plasmid encoding MbPylRS(IPYE) and pylT. ACCESSION . VERSION . KEYWORDS pTECH-MbPylRS(IPYE) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3839) AUTHORS Bryson DI, Fan C, Guo LT, Miller C, Soll D, Liu DR TITLE Continuous directed evolution of aminoacyl-tRNA synthetases. JOURNAL Nat Chem Biol. 2017 Oct 16. doi: 10.1038/nchembio.2474. PUBMED 29035361 REFERENCE 2 (bases 1 to 3839) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3839 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(2513..2541) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" rep_origin 2653..3198 /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 3099..3118 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3315..3332 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(3687..3706) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" ORIGIN 1 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc 61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc 121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggcgaaaa cctggcccta 181 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt 241 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat 301 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca 361 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga 421 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg 481 ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga aagcaaattc 541 gacccgggcg gccgcaaccc agcgttcgat gcttctttga gcgaacgatc aaaaataagt 601 gccttcccat caaaaaaata ttctcaacat aaaaaacttt gtgtaatact tgtaacgcta 661 catggagatt aactcaatct agagggtaac tagtatggat aagaagccgc tggatgttct 721 gatctctgcg accggtctgt ggatgtcccg taccggcacg ctgcacaaga tcaagcacta 781 tgagatttct cgttctaaaa tctacatcga aatggcgtgt ggtgaccatc tggttgtgaa 841 caactctcgt tcttgtcgtc ccgcacgtgc attccgttat cataaatacc gtaaaacctg 901 caaacgttgt cgtgtttctg acgaagatat caacaacttc ctgacccgtt ctaccgaagg 961 caaaacctct gttaaagtta aagttgtttc tgagccgaaa gtgaaaaaag cgatgccgaa 1021 atctgtttct cgtgcgccga aaccgctgga aaatccggtt tctgcgaaag cgtctaccga 1081 cacctctcgt tctgttccgt ctccggcgaa atctaccccg aactctccgg ttccgacctc 1141 tgcgccggcg ccgtctctga cccgttctca gctggatcgt gttgaagcgc tgctgtctcc 1201 ggaagataaa atctctctga acatcgcgaa accgttccgt gaactggaat ctgaactggt 1261 tacccgtcgt aaaaacgatt tccagcgtct gtacaccaac gatcgtgaag actacctggg 1321 taaactggaa cgtgacatca ccaaattctt cgttgaccgt gatttcctgg aaatcaaatc 1381 tccgatcctg atcccggcgg aatacgttga acgtatgggt atcaacaacg ataccgaact 1441 gtctaaacag atcttccgtg ttgataaaaa cctgtgcctg cgtccgatgc tggcgccgac 1501 cctgtacaac tatctgcgta aactggatcg tatcctgccg gacccgatca aaatcttcga 1561 agttggtccg tgctaccgta aagaatctga cggtaaagaa cacctggaag agttcaccat 1621 ggtgaacttc tgccagatgg gttctggttg cacccgtgag aacctggaat ctctgatcaa 1681 agaatttctg gactacctgg aaatcgactt cgaaatcgtt ggtgactcct gcatggtgta 1741 cggtgatacc ctggacatca tgcacggtga cctggaactg tcttctgcgg ttgttggtcc 1801 ggttccgctg gatcgtgaat ggggtatcga caaaccgtgg atcggtgcgg gtttcggtct 1861 ggaacgtctg ctgaaagtta tgcacggttt caaaaacatc aaacgtgcgt ctcgttctga 1921 atcttactac aacggtatct ctaccaacct gtaacctagg ctgctgccac cgctgagatc 1981 tatgccccgc gcccacagga aggagctacc ggcagcggtg cggactgttg taactcataa 2041 taagaaatga ggccgctcat ggcgttctgt tgcccgtctc gctggtgaaa agaaaaacca 2101 ccctggctaa ctaagcggcc tgctgacttt ctcgccgatc aaaaggcatt ttgctattaa 2161 gggattgacg agggcgtatc tgcgcagtaa gatgcgcccc gcattggaaa cctgatcatg 2221 tagatcgaat ggactctaaa tccgttcagc cgggttagat tcccggggtt tccgccaaat 2281 tcgaaaagcc tgctcaacga gcaggctttt ttgcatggcg ttacccaact taatcgcctt 2341 gcagcacatc cccctttcgc cagatcccgc aagaggcccg gcagtaccgg cataaccaag 2401 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat 2461 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc 2521 ttatcgatga taagctgtca aacatgagaa ttacaactta tatcgtatgg ggctgacttc 2581 aggtgctaca tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat 2641 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa 2701 aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg 2761 taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc 2821 gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt 2881 ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 2941 actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga 3001 gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga 3061 aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat 3121 agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg 3181 gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct 3241 tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc 3301 gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat 3361 tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc 3421 ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg 3481 cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagta agaggttcca 3541 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt 3601 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat 3661 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata 3721 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca 3781 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaactc //