LOCUS       sequence-323479-        6699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose-inducible expression of six mutagenesis genes, each 
            encoding one amber codon after the initiator methionine.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6699)
  AUTHORS   Ho JML, Miller CA, Parks SE, Mattia JR, Bennett MR
  TITLE     A suppressor tRNA-mediated feedforward loop eliminates leaky gene 
            expression in bacteria.
  JOURNAL   Nucleic Acids Res. 2021 Mar 18;49(5):e25. doi: 10.1093/nar/gkaa1179.
  PUBMED    33290521
REFERENCE   2  (bases 1 to 6699)
  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: "Nucleic 
            Acids Res."; date: "2021-03-18"; volume: "49(5):e25. doi"; pages: " 
            10.1093/nar/gkaa1179"
COMMENT     Sequence Label: 
            MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG
FEATURES             Location/Qualifiers
     source          1..6699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(39..777)
                     /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."
     terminator      867..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(927..1805)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1832..2116
                     /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     2039..2058
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4936..5193
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="M*TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVH
                     TAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5223..5852
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="M*TDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGE
                     RRACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEK
                     ILEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKI
                     FIQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     terminator      complement(5879..5910)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(5926..6585)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
       61 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
      121 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
      181 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
      241 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
      301 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
      361 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
      421 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
      481 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
      541 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
      601 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
      661 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
      721 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      781 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
      841 agcccaggag gaagaggaca tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg
      901 cctttcgttt tagacttagg gaccctttat gacaacttga cggctacatc attcactttt
      961 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
     1021 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
     1081 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
     1141 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
     1201 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
     1261 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
     1321 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
     1381 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
     1441 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
     1501 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
     1561 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
     1621 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
     1681 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
     1741 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     1801 gccatacttt tcatactccc accattcaga gaagaaacca attgtccata ttgcatcaga
     1861 cattgccgtc actgcgtctt ttactggctc ttctcgctaa cccaaccggt aaccccgctt
     1921 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
     1981 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
     2041 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
     2101 tctactgttt ctccataccc gtttttttgg acgcgtacaa ctcaagtctg acataaatga
     2161 ccgctatgta gagcactgca attacacgcc agatcgttct cgctaccgca accaccggta
     2221 tgaaccagat tggtgcgcac tatgaaggcc acaagatcat tgagattggt gccgttgaag
     2281 tggtgaaccg tcgcctgacg ggcaataact tccatgttta tctcaaaccc gatcggctgg
     2341 tggatccgga agcctttggc gtacatggta ttgccgatga atttttgctc gataagccca
     2401 cgtttgccga agtagccgat gagttcatgg actatattcg cggcgcggag ttggtgatcc
     2461 ataacgcagc gttcgatatc ggctttatgg actacgagtt ttcgttgctt aagcgcgata
     2521 ttccgaagac caatactttc tgtaaggtca ccgatagcct tgcggtggcg aggaaaatgt
     2581 ttcccggtaa gcgcaacagc ctcgatgcgt tatgtgctcg ctacgaaata gataacagta
     2641 aacgaacgct gcacggggca ttactcgatg cccagatcct tgcggaagtt tatctggcga
     2701 tgaccggtgg tcaaacgtcg atggcttttg cgatggaagg agagacacaa cagcaacaag
     2761 gtgaagcaac aattcagcgc attgtacgtc aggcaagtaa gttacgcgtt gtttttgcga
     2821 cagatgaaga gattgcagct catgaagccc gtctcgatct ggtgcagaag aaaggcggaa
     2881 gttgcctctg gcgagcataa tttaatatca gtaaaccgga cataacccat gtagaagaaa
     2941 aatcgcgctt ttttgaagtg ggcagggggc aagtatcccc tgcttgatga tattaaacgg
     3001 catttgccca agggcgaatg tctggttgag ccttttgtag gtgccgggtc ggtgtttctc
     3061 aacaccgact tttctcgtta tatccttgcc gatatcaata gcgacctgat cagtctctat
     3121 aacattgtga agatgcgtac tgatgagtac gtacaggccg cacgcgagct gtttgttccc
     3181 gaaacaaatt gcgccgaggt ttactatcag ttccgcgaag agttcaacaa aagccaggat
     3241 ccgttccgtc gggcggtact gtttttatat ttgaaccgct acggttacaa cggcctgtgt
     3301 cgttacaatc tgcgcggtga gtttaacgtg ccgttcggcc gctacaaaaa accctatttc
     3361 ccggaagcag agttgtatca cttcgctgaa aaagcgcaga atgccttttt ctattgtgag
     3421 tcttacgccg atagcatggc gcgcgcagat gatgcatccg tcgtctattg cgatccgcct
     3481 tatgcaccgc tgtctgcgac cgccaacttt acggcgtatc acacaaacag ttttacgctt
     3541 gaacaacaag cgcatctggc ggagatcgcc gaaggtctgg ttgagcgcca tattccagtg
     3601 ctgatctcca atcacgatac gatgttaacg cgtgagtggt atcagcgcgc aaaattgcat
     3661 gtcgtcaaag ttcgacgcag tataagcagc aacggcggca cacgtaaaaa ggtggacgaa
     3721 ctgctggctt tgtacaaacc aggagtcgtt tcacccgcga aaaaataatt cagctaagac
     3781 actgcactgg attaagatgt agaaaacgat tgaagttgat gatgaactct acagctatat
     3841 tgccagccac actaagcata tcggcgagag cgcatccgac attttacggc gtatgttgaa
     3901 attttccgcc gcatcacagc ctgctgctcc ggtgacgaaa gaggttcgcg ttgcgtcacc
     3961 tgctatcgtc gaagcgaagc cggtcaaaac gattaaagac aaggttcgcg caatgcgtga
     4021 acttctgctt tcggatgaat acgcagagca aaagcgagcg gtcaatcgct ttatgctgct
     4081 gttgtctaca ctatattctc ttgacgccca ggcgtttgcc gaagcaacgg aatcgttgca
     4141 cggtcgtaca cgcgtttact ttgcggcaga tgaacaaacg ctgctgaaaa atggtaatca
     4201 gaccaagccg aaacatgtgc caggcacgcc gtattgggtg atcaccaaca ccaacaccgg
     4261 ccgtaaatgc agcatgatcg aacacatcat gcagtcgatg caattcccgg cggaattgat
     4321 tgagaaggtt tgcggaacta tctaacggct gaaattaatg aggtcatacc caaatgtagg
     4381 atagttcgtt tacgcccatt gaacaaatgc taaaatttcg cgccagccgc cacgaagatt
     4441 ttccttatca ggagatcctt ctgactcgtc tttgcatgca catgcaaagc aagctgctgg
     4501 agaaccgcaa taaaatgctg aaggctcagg gaattaacga gacgttgttt atggcgttga
     4561 ttacgctgga gtctcaggaa aaccacagta ttcagccttc tgaattaagt tgtgctcttg
     4621 gatcatcccg taccaacgcg acgcgtattg ccgatgaact ggaaaaacgc ggttggatcg
     4681 aacgtcgtga aagcgataac gatcgccgct gcctgcatct gcaattaacg gaaaaaggtc
     4741 acgagttttt gcgcgaggtt ttaccaccgc agcataactg cctgcatcaa ctctggtccg
     4801 cgctcagcac aacagaaaaa gatcagctcg agcaaatcac ccgcaaattg ctctcccgtc
     4861 tcgaccagat ggaacaagac ggtgtggttc tcgaagcgat gagctaataa tacaaaaatt
     4921 aggaggaatt tcaacatgta gacaaattta tctgacatca ttgaaaaaga aacaggaaaa
     4981 caactagtga ttcaagaatc aattctaatg ttaccagaag aagtagagga agtaattggg
     5041 aataaaccag aaagtgatat tttagttcat actgcttatg atgaaagtac agatgaaaat
     5101 gtaatgctat taacttcaga tgctccagaa tataaacctt gggctttagt aattcaagac
     5161 agtaatggag aaaataaaat taaaatgtta taagtcgaga ttaagtaaac cggaatctga
     5221 agatgtagac cgacgcggaa tacgttcgta tccacgaaaa actggacatc tacaccttca
     5281 aaaaacagtt cttcaacaac aaaaaatctg tttctcaccg ttgctacgtt ctgttcgaac
     5341 tgaaacgtcg tggtgaacgt cgtgcgtgct tctggggtta cgcggttaac aaaccgcagt
     5401 ctggtaccga acgtggtatc cacgcggaaa tcttctctat ccgtaaagtt gaagaatacc
     5461 tgcgtgacaa cccgggtcag ttcaccatca actggtactc ttcttggtct ccgtgcgcgg
     5521 actgcgcgga aaaaatcctg gaatggtaca accaggaact gcgtggtaac ggtcacaccc
     5581 tgaaaatctg ggcgtgcaaa ctgtactacg aaaaaaacgc gcgtaaccag atcggtctgt
     5641 ggaacctgcg tgacaacggt gttggtctga acgttatggt ttctgaacac taccagtgct
     5701 gccgtaaaat cttcatccag tcttctcaca accagctgaa cgaaaaccgt tggctggaaa
     5761 aaaccctgaa acgtgcggaa aaacgtcgtt ctgaactgtc tatcatgatc caggttaaaa
     5821 tcctgcacac caccaaatct ccggcggttt aaacttaatt aacggcactc ctcagccaag
     5881 tcaaaagcct ccggtcggag gcttttgact acatgcccat ggcgtttacg ccccgccctg
     5941 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     6001 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     6061 tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     6121 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     6181 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     6241 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     6301 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     6361 catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     6421 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     6481 ctggttatag gtacattgag taactgactg aaatgcctca aaatgttctt tacgatgcca
     6541 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     6601 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     6661 aaagttggaa cctcttacgt gccaagccaa ataggccgt
//
