LOCUS Exported 5652 bp ds-DNA circular SYN 19-APR-2022 DEFINITION N‐acetyl lysyl‐tRNA synthetase with cognate tRNA for genetic code expansion with ochre suppression. ACCESSION . VERSION . KEYWORDS pEVOL-MmAcKRS1-PylTUUA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5652) AUTHORS Wu B, Wang Z, Huang Y, Liu WR TITLE Catalyst-free and site-specific one-pot dual-labeling of a protein directed by two genetically incorporated noncanonical amino acids. JOURNAL Chembiochem. 2012 Jul 9;13(10):1405-8. doi: 10.1002/cbic.201200281. Epub 2012 May 24. PUBMED 22628069 REFERENCE 2 (bases 1 to 5652) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 19, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5652 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(480..497) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(614..1159) /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(694..713) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1305..2183) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 2210..2494 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 2417..2436 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 3902..3919 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(3975..3992) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" CDS complement(4978..5637) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 5552..5571 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(5638..88) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" ORIGIN 1 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg 61 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg 121 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac 181 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat 241 gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag 301 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg 361 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg 421 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc 481 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa 541 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg 601 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct 661 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg 721 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat 781 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag 841 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat 901 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat 961 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag 1021 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag 1081 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg 1141 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc 1201 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta 1261 attctcatgt ttgacagctt atcatcgata agcttggtac ccaattatga caacttgacg 1321 gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg 1381 cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg 1441 gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc 1501 tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac 1561 ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga 1621 tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg 1681 ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc 1741 gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc 1801 ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta 1861 agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga 1921 gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcactc 1981 ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga 2041 ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg 2101 gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg 2161 ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat 2221 tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc 2281 aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca 2341 aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac 2401 ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga 2461 cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc taacaggagg 2521 aattactagt atggataaga aaccgctgaa tactctgatt tctgcaactg gtctgtggat 2581 gagccatacc ggcaccatcc acaagatcaa acaccgcgag gtttcccgaa gcaaaatcta 2641 catcgaaatg gcgtgcggtg accacctggt ggtaaacaac tcccgttctt ctcgtactgc 2701 acgtgctctg cgccaccaca agtaccgtaa gacctgcaag cgctgtcgcg tgtctgatga 2761 agacctgaac aaattcctga ctaaagcgaa cgaagatcag acttctgtga aggtgaaagt 2821 tgtttctgcc ccaacccgca ccaagaaagc gatgccgaag tccgttgcac gcgctccgaa 2881 accgctggag aactccgaag ccgcacaggc ccagccgtct ggttctaagt tttctccggc 2941 aatcccggtt tctactcagg agtctgtgtc tgtgccagct tctgttagca cttctatttc 3001 ctctatcagc actggtgcga ctgcgtccgc tctggtaaaa ggtaacacta acccgatcac 3061 cagcatgtct gctccggttc aggcttctgc accggcactg actaaaagcc agactgaccg 3121 tctggaggtt ctgctgaacc cgaaagatga aatcagcctg aactctggca aaccgttccg 3181 tgaactggaa tccgaactgc tgtctcgtcg taagaaagac ctgcaacaaa tctatgctga 3241 agagcgtgaa aactacctgg gtaaactgga acgtgaaatc acccgtttct ttgtggaccg 3301 tggtttcctg gaaatcaagt ctccgatcct gatcccgctg gaatacatcg agcgcatggg 3361 tattgataac gacaccgaac tgtccaagca gattttccgt gtggacaaga acttctgcct 3421 gcgtccgatg atggcaccga acctgctgaa ttacgcacgt aaactggatc gtgcactgcc 3481 ggacccgatc aaaatctttg aaatcggtcc atgctatcgt aaggagagcg acggtaaaga 3541 acacctggaa gagttcacta tgctgaactt ttttcagatg ggttctggct gcacccgtga 3601 aaatctggaa tctatcatca ccgacttcct gaaccacctg ggcattgact tcaaaatcgt 3661 tggtgattcc tgcatggttt acggtgacac tctggacgtt atgcatggtg atctggaact 3721 gagcagcgct gttgtgggtc cgattccgct ggatcgtgaa tggggtatcg ataaaccgtg 3781 gattggtgct ggcttcggtc tggaacgtct gctgaaagtt aagcacgact ttaagaacat 3841 caaacgtgct gcgcgttccg agtcctatta caacggcatt agcactaacc tgtaagtcga 3901 ccatcatcat catcatcatt gagtttaaac ggtctccagc ttggctgttt tggcggatga 3961 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag 4021 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgtcgaa ctcagaagtg 4081 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag 4141 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc ttgtttgtga gctcccggtc 4201 atcaatcatc cccataatcc ttgttagatt atcaatttta aaaaactaac agttgtcagc 4261 ctgtcccgct ttaatatcat acgccgttat acgttgttta cgctttgagg aatcccatat 4321 gatacgcggc cgctttcaaa cgctaaattg cctgatgcgc tacgcttatc aggcctacat 4381 gatctctgca atatattgag tttgcgtgct tttgtaggcc ggataaggcg ttcacgccgc 4441 atccggcaag aaacagcaaa caatccaaaa cgccgcgttc agcgacgttt tttctgcttt 4501 tcttcgcgaa ttaattccgc ttcgcaacat gtgagcaccg gtttattgac taccggaagc 4561 agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct ccccgtggag 4621 gtaataattg acgatatgat cagtgcacgg ctaactaagc ggcctgctga ctttctcgcc 4681 gatcaaaagg cattttgcta ttaagggatt gacgagggcg tatctgcgca gtaagatgcg 4741 ccccgcattg gaaacctgat catgtagatc gaatggactt taaatccgtt cagccgggtt 4801 agattcccgg ggtttccgcc aaattcgaaa agcctgctca acgagcaggc ttttttgcat 4861 gctcgagcag ctcagggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc 4921 acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta 4981 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg 5041 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc 5101 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac 5161 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc 5221 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata 5281 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc 5341 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc 5401 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat 5461 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc 5521 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc 5581 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt 5641 agcttcctta gc //