LOCUS       sequence-250213-        5361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Double mutant of wild type mmPylRS designed for incorporation of 
            non-cannonical amino acids in to M13 bacteriophage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5361)
  AUTHORS   Tharp JM, Hampton JT, Reed CA, Ehnbom A, Chen PC, Morse JS, Kurra Y,
            Perez LM, Xu S, Liu WR
  TITLE     An amber obligate active site-directed ligand evolution technique 
            for phage display.
  JOURNAL   Nat Commun. 2020 Mar 13;11(1):1392. doi: 10.1038/s41467-020-15057-7.
  PUBMED    32170178
REFERENCE   2  (bases 1 to 5361)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-03-13"; volume: "11(1):1392. doi"; pages: " 
            10.1038/s41467-020-15057-7"
COMMENT     Sequence Label: pEDF-PhdRS
FEATURES             Location/Qualifiers
     source          1..5361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
//
