LOCUS       sequence-243924-        7077 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 7077)
  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 7077)
  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..7077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(490..1228)
                     /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(1365..2243)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2270..2554
                     /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     2477..2496
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3962..3979
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4035..4052)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(6403..7062)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6977..6996
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(7063..7077,1..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 ggcgcgccca aatagctagc tcactcggtc gctacgctcc gggcgtgaga
      481 ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg
      541 actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc
      601 ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc
      661 tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt
      721 ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg
      781 ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct
      841 cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc
      901 cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa
      961 aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac
     1021 acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt
     1081 gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc
     1141 gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa
     1201 aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctctaga tttcagtgca
     1261 atttatccct aggtcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     1321 attctcatgt ttgacagctt atcatcgata agcttggtac ccaattatga caacttgacg
     1381 gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg
     1441 cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg
     1501 gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc
     1561 tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac
     1621 ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga
     1681 tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg
     1741 ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc
     1801 gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc
     1861 ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta
     1921 agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga
     1981 gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcactc
     2041 ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga
     2101 ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg
     2161 gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg
     2221 ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat
     2281 tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc
     2341 aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca
     2401 aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac
     2461 ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga
     2521 cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc taacaggagg
     2581 aattactagt atggataaaa aaccactaaa cactctgata tctgcaaccg ggctctggat
     2641 gtccaggacc ggaacaattc ataaaataaa acaccacgaa gtctctcgaa gcaaaatcta
     2701 tattgaaatg gcatgcggag accaccttgt tgtaaacaac tccaggagca gcaggactgc
     2761 aagagcgctc aggcaccaca aatacaggaa gacctgcaaa cgctgcaggg tttcggatga
     2821 ggatctcaat aagttcctca caaaggcaaa cgaagaccag acaagcgtaa aagtcaaggt
     2881 cgtttctgcc cctaccagaa cgaaaaaggc aatgccaaaa tccgttgcga gagccccgaa
     2941 acctcttgag aatacagaag cggcacaggc tcaaccttct ggatctaaat tttcacctgc
     3001 gataccggtt tccacccaag agtcagtttc tgtcccggca tctgtttcaa catcaatatc
     3061 aagcatttct acaggagcaa ctgcatccgc actggtaaaa gggaatacga accccattac
     3121 atccatgtct gcccctgttc aggcaagtgc ccccgcactt acgaagagcc agactgacag
     3181 gcttgaagtc ctgttaaacc caaaagatga gatttccctg aattccggca agcctttcag
     3241 ggagcttgag tccgaattgc tctctcgcag aaaaaaagac ctgcagcaga tctacgcgga
     3301 agaaagggag aattatctgg ggaaactcga gcgtgaaatt accaggttct ttgtggacag
     3361 gggttttctg gaaataaaat ccccgatcct gatccctctt gagtatatcg aaaggatggg
     3421 cattgataat gataccgaac tttcaaaaca gatcttcagg gttgacaaga acttctgcct
     3481 gagacccatg cttgctccaa acctttacaa ctacctgcgc aagcttgaca gggccctgcc
     3541 tgatccaata aaaatttttg aaataggccc atgctacaga aaagagtccg acggcaaaga
     3601 acacctcgaa gagtttacca tgctggcgtt cgcgcagatg ggatcgggat gcacacggga
     3661 aaatcttgaa agcataatta cggacttcct gaaccacctg ggaattgatt tcaagatcgt
     3721 aggcgattcc tgcatggtct atggggatac ccttgatgta atgcacggag acctggaact
     3781 ttcctctgca gtagtcggac ccataccgct tgaccgggaa tggggtattg ataaaccctg
     3841 gataggggca ggtttcgggc tcgaacgcct tctaaaggtt aaacacgact ttaaaaatat
     3901 caagagagct gcaaggtccg agtcttacta taacgggatt tctaccaacc tgtaagtcga
     3961 ccatcatcat catcatcatt gagtttaaac ggtctccagc ttggctgttt tggcggatga
     4021 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     4081 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     4141 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     4201 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc ttgtttgtga gctcccggtc
     4261 atcaatcatc cccataatcc ttgttagatt atcaatttta aaaaactaac agttgtcagc
     4321 ctgtcccgct ttaatatcat acgccgttat acgttgttta cgctttgagg aatcccatat
     4381 gatggataaa aaaccactaa acactctgat atctgcaacc gggctctgga tgtccaggac
     4441 cggaacaatt cataaaataa aacaccacga agtctctcga agcaaaatct atattgaaat
     4501 ggcatgcgga gaccaccttg ttgtaaacaa ctccaggagc agcaggactg caagagcgct
     4561 caggcaccac aaatacagga agacctgcaa acgctgcagg gtttcggatg aggatctcaa
     4621 taagttcctc acaaaggcaa acgaagacca gacaagcgta aaagtcaagg tcgtttctgc
     4681 ccctaccaga acgaaaaagg caatgccaaa atccgttgcg agagccccga aacctcttga
     4741 gaatacagaa gcggcacagg ctcaaccttc tggatctaaa ttttcacctg cgataccggt
     4801 ttccacccaa gagtcagttt ctgtcccggc atctgtttca acatcaatat caagcatttc
     4861 tacaggagca actgcatccg cactggtaaa agggaatacg aaccccatta catccatgtc
     4921 tgcccctgtt caggcaagtg cccccgcact tacgaagagc cagactgaca ggcttgaagt
     4981 cctgttaaac ccaaaagatg agatttccct gaattccggc aagcctttca gggagcttga
     5041 gtccgaattg ctctctcgca gaaaaaaaga cctgcagcag atctacgcgg aagaaaggga
     5101 gaattatctg gggaaactcg agcgtgaaat taccaggttc tttgtggaca ggggttttct
     5161 ggaaataaaa tccccgatcc tgatccctct tgagtatatc gaaaggatgg gcattgataa
     5221 tgataccgaa ctttcaaaac agatcttcag ggttgacaag aacttctgcc tgagacccat
     5281 gcttgctcca aacctttaca actacctgcg caagcttgac agggccctgc ctgatccaat
     5341 aaaaattttt gaaataggcc catgctacag aaaagagtcc gacggcaaag aacacctcga
     5401 agagtttacc atgctggcgt tcgcgcagat gggatcggga tgcacacggg aaaatcttga
     5461 aagcataatt acggacttcc tgaaccacct gggaattgat ttcaagatcg taggcgattc
     5521 ctgcatggtc tatggggata cccttgatgt aatgcacgga gacctggaac tttcctctgc
     5581 agtagtcgga cccataccgc ttgaccggga atggggtatt gataaaccct ggataggggc
     5641 aggtttcggg ctcgaacgcc ttctaaaggt taaacacgac tttaaaaata tcaagagagc
     5701 tgcaaggtcc gagtcttact ataacgggat ttctaccaac ctgtaagcgg ccgctttcaa
     5761 acgctaaatt gcctgatgcg ctacgcttat caggcctaca tgatctctgc aatatattga
     5821 gtttgcgtgc ttttgtaggc cggataaggc gttcacgccg catccggcaa gaaacagcaa
     5881 acaatccaaa acgccgcgtt cagcggcgtt ttttctgctt ttcttcgcga attaattccg
     5941 cttcgcaaca tgtgagcacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct
     6001 caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga
     6061 tcagtgcacg gctaactaag cggcctgctg actttctcgc cgatcaaaag gcattttgct
     6121 attaagggat tgacgagggc gtatctgcgc agtaagatgc gccccgcatt ccggcggtag
     6181 ttcagcaggg cagaacggcg gactctaaat ccgcatggca ggggttcaaa tcccctccgc
     6241 cggaccaaat tcgaaaagcc tgctcaacga gcaggctttt ttgcatgctc gagcagctca
     6301 gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt
     6361 agcaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca
     6421 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacagac
     6481 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     6541 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     6601 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     6661 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     6721 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     6781 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     6841 acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     6901 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     6961 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     7021 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagc
//
