LOCUS Exported 5361 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Double mutant of wild type mmPylRS designed for incorporation of non-cannonical amino acids in to M13 bacteriophage. ACCESSION . VERSION . KEYWORDS pEDF-PhdRS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5361) TITLE An Amber-Codon-Obligate Phage Display System to Assist Drug Discovery JOURNAL Unpublished REFERENCE 2 (bases 1 to 5361) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5361 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..285 /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 208..227 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 1693..1710 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1766..1783) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" CDS complement(2418..3077) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 2992..3011 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(3078..3180) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" rep_origin complement(3582..4320) /direction=LEFT /label=CloDF13 ori /note="Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." CDS complement(4457..5335) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" ORIGIN 1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct 61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca 121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg 181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg 241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg 301 ctaacaggag gaattactag tatggataaa aaaccactaa acactctgat atctgcaacc 361 gggctctgga tgtccaggac cggaacaatt cataaaataa aacaccacga agtctctcga 421 agcaaaatct atattgaaat ggcatgcggt gaccaccttg ttgtaaacaa ctccaggagc 481 agcaggactg caagagcgct caggcaccac aaatacagga agacctgcaa acgctgcagg 541 gtttcggatg aggatctcaa taagttcctc acaaaggcaa acgaagacca gacaagcgta 601 aaagtcaagg tcgtttctgc ccctaccaga acgaaaaagg caatgccaaa atccgttgcg 661 agagccccga aacctcttga gaatacagaa gcggcacagg ctcaaccttc tggatctaaa 721 ttttcacctg cgataccggt ttccacccaa gagtcagttt ctgtcccggc atctgtttca 781 acatcaatat caagcatttc tacaggagca actgcatccg cactggtaaa agggaatacg 841 aaccccatta catccatgtc tgcccctgtt caggcaagtg cccccgcact tacgaagagc 901 cagactgaca ggcttgaagt cctgttaaac ccaaaagatg agatttccct gaattccggc 961 aagcctttca gggagcttga gtccgaattg ctctctcgca gaaaaaaaga cctgcagcag 1021 atctacgcgg aagaaaggga gaattatctg gggaaactcg agcgtgaaat taccaggttc 1081 tttgtggaca ggggttttct ggaaataaaa tccccgatcc tgatccctct tgagtatatc 1141 gaaaggatgg gcattgataa tgataccgaa ctttcaaaac agatcttcag ggttgacaag 1201 aacttctgcc tgagacccat gcttgctcca aacctttaca actacctgcg caagcttgac 1261 agggccctgc ctgatccaat aaaaattttt gaaataggcc catgctacag aaaagagtcc 1321 gacggcaaag aacacctcga agagtttacc atgctggcgt tcgcgcagat gggatcggga 1381 tgcacacggg aaaatcttga aagcataatt acggacttcc tgaaccacct gggaattgat 1441 ttcaagatcg taggcgattc ctgcatggtc tatggggata cccttgatgt aatgcacgga 1501 gacctggaac tttcctctgc agtagtcgga cccataccgc ttgaccggga atggggtatt 1561 gataaaccct ggataggggc aggtttcggg ctcgaacgcc ttctaaaggt taaacacgac 1621 tttaaaaata tcaagagagc tgcaaggtcc gagtcttact ataacgggat ttctaccaac 1681 ctgtaagtcg accatcatca tcatcatcat tgagtttaaa cggtctccag cttggctgtt 1741 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc 1801 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga 1861 actcagaagt gaaacgccgc atccggcaag aaacagcaaa caatccaaaa cgccgcgttc 1921 agcggcgttt tttctgcttt tcttcgcgaa ttaattccgc ttcgcaacat gtgagcaccg 1981 gtttattgac taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg 2041 ctcaggctct ccccgtggag gtaataattg acgatatgat cagtgcacgg ctaactaagc 2101 ggcctgctga ctttctcgcc gatcaaaagg cattttgcta ttaagggatt gacgagggcg 2161 tatctgcgca gtaagatgcg ccccgcattg gaaacctgat catgtagatc gaatggactc 2221 taaatccgtt cagccgggtt agattcccgg ggtttccgcc aaattcgaaa agcctgctca 2281 acgagcaggc ttttttgcat gctcgagcag ctcagggtcg aatttgcttt cgaatttctg 2341 ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa 2401 taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca 2461 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc 2521 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag 2581 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct 2641 gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa 2701 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc 2761 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta 2821 tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg agcattcatc 2881 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc 2941 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac 3001 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca 3061 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat 3121 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca 3181 acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga 3241 tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca aagtgcgtcg 3301 ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg ctccagtggc 3361 ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa 3421 aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat 3481 gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggggcgcgcc caaatagcta 3541 gctcactcgg tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa 3601 agttacccac agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc 3661 cgcgccggtg gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg 3721 cttttccggt gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga 3781 aacccgacag gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc 3841 ctgttccgac cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt 3901 ggcgctttct catagctcac acactggtat ctcggctcgg tgtaggtcgt tcgctccaag 3961 ctgggctgta agcaagaact ccccgttcag cccgactgct gcgccttatc cggtaactgt 4021 tcacttgagt ccaacccgga aaagcacggt aaaacgccac tggcagcagc cattggtaac 4081 tgggagttcg cagaggattt gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa 4141 agtccggcta cactggaagg acagatttgg ttgctgtgct ctgcgaaagc cagttaccac 4201 ggttaagcag ttccccaact gacttaacct tcgatcaaac cacctcccca ggtggttttt 4261 tcgtttacag ggcaaaagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 4321 ttttctactg aaccgctcta gatttcagtg caatttatcc ctaggtcttc aaatgtagca 4381 cctgaagtca gccccatacg atataagttg taattctcat gtttgacagc ttatcatcga 4441 taagcttggt acccaattat gacaacttga cggctacatc attcactttt tcttcacaac 4501 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga 4561 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca 4621 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta 4681 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc 4741 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta 4801 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca 4861 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt 4921 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga 4981 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac 5041 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa 5101 tctctcctgg cgggaacagc aaaatatcac tcggtcggca aacaaattct cgtccctgat 5161 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg 5221 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca 5281 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt 5341 tcatactccc gccattcaga g //