LOCUS       sequence-368421-        8550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eMutaT7[TadA-8e] for mutagenizing genes downstream from a 
            T7 promoter in e.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8550)
  AUTHORS   Seo D, Koh B, Eom GE, Kim HW, Kim S
  TITLE     A dual gene-specific mutator system installs all transition 
            mutations at similar frequencies in vivo.
  JOURNAL   Nucleic Acids Res. 2023 Apr 18:gkad266. doi: 10.1093/nar/gkad266.
  PUBMED    37070179
REFERENCE   2  (bases 1 to 8550)
  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. 2023 Apr 18:gkad266. doi: 10.1093/nar/gkad266."
COMMENT     Sequence Label: pDae029 (eMutaT7[TadA-8e])
FEATURES             Location/Qualifiers
     source          1..8550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(4584..4601)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4754..4840
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4932..4959
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4978..5069
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5527..5982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5614..5633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5824..5845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6538..7197)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7112..7131
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7198..7300)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(7692..7709)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7826..8371)
                     /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(7906..7925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8447..8466
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcgaatt cgagctcagg
     1321 aggaattcac catggcatat gagtgaagtt gaattcagcc atgaatattg gatgcgtcat
     1381 gccctgaccc tggcaaaacg tgcccgcgat gaacgtgaag tgccggttgg tgcggtgctg
     1441 gttttaaata atcgtgtgat tggcgaaggc tggaatcgtg cgattggtct gcatgatcct
     1501 accgcacatg cagaaatcat ggccctgcgt cagggtggtc tggttatgca aaattatcgc
     1561 ctgattgatg ccaccctgta tgtgaccttt gaaccttgtg tgatgtgtgc gggtgcaatg
     1621 attcacagtc gtattggccg tgttgtgttt ggcgttcgta atagcaagcg cggtgccgcc
     1681 ggcagcctga tgaatgttct gaattatcct ggcatgaatc atcgcgtgga aattaccgag
     1741 ggtattctgg ccgatgaatg tgccgcactg ctgtgcgatt tttatcgcat gccgcgtcaa
     1801 gtgtttaatg cacaaaagaa agcacagagc agcattaata gcggcagcga gactcccggg
     1861 acctcagagt ccgccacacc cgaaagtaac acgattaaca tcgctaagaa cgacttctct
     1921 gacatcgaac tggctgctat cccgttcaac actctggctg accattacgg tgagcgttta
     1981 gctcgcgaac agttggccct tgagcatgag tcttacgaga tgggtgaagc acgcttccgc
     2041 aagatgtttg agcgtcaact taaagctggt gaggttgcgg ataacgctgc cgccaagcct
     2101 ctcatcacta ccctactccc taagatgatt gcacgcatca acgactggtt tgaggaagtg
     2161 aaagctaagc gcggcaagcg cccgacagcc ttccagttcc tgcaagaaat caagccggaa
     2221 gccgtagcgt acatcaccat taagaccact ctggcttgcc taaccagtgc tgacaataca
     2281 accgttcagg ctgtagcaag cgcaatcggt cgggccattg aggacgaggc tcgcttcggt
     2341 cgtatccgtg accttgaagc taagcacttc aagaaaaacg ttgaggaaca actcaacaag
     2401 cgcgtagggc acgtctacaa gaaagcattt atgcaagttg tcgaggctga catgctctct
     2461 aagggtctac tcggtggcga ggcgtggtct tcgtggcata aggaagactc tattcatgta
     2521 ggagtacgct gcatcgagat gctcattgag tcaaccggaa tggttagctt acaccgccaa
     2581 aatgctggcg tagtaggtca agactctgag actatcgaac tcgcacctga atacgctgag
     2641 gctatcgcaa cccgtgcagg tgcgctggct ggcatctctc cgatgttcca accttgcgta
     2701 gttcctccta agccgtggac tggcattact ggtggtggct attgggctaa cggtcgtcgt
     2761 cctctggcgc tggtgcgtac tcacagtaag aaagcactga tgcgctacga agacgtttac
     2821 atgcctgagg tgtacaaagc gattaacatt gcgcaaaaca ccgcatggaa aatcaacaag
     2881 aaagtcctag cggtcgccaa cgtaatcacc aagtggaagc attgtccggt cgaggacatc
     2941 cctgcgattg agcgtgaaga actcccgatg aaaccggaag acatcgacat gaatcctgag
     3001 gctctcaccg cgtggaaacg tgctgccgct gctgtgtacc gcaaggacaa ggctcgcaag
     3061 tctcgccgta tcagccttga gttcatgctt gagcaagcca ataagtttgc taaccataag
     3121 gccatctggt tcccttacaa catggactgg cgcggtcgtg tttacgctgt gtcaatgttc
     3181 aacccgcaag gtaacgatat gaccaaagga ctgcttacgc tggcgaaagg taaaccaatc
     3241 ggtaaggaag gttactactg gctgaaaatc cacggtgcaa actgtgcggg tgtcgataag
     3301 gttccgttcc ctgagcgcat caagttcatt gaggaaaacc acgagaacat catggcttgc
     3361 gctaagtctc cactggagaa cacttggtgg gctgagcaag attctccgtt ctgcttcctt
     3421 gcgttctgct ttgagtacgc tggggtacag caccacggcc tgagctataa ctgctccctt
     3481 ccgctggcgt ttgacgggtc ttgctctggc atccagcact tctccgcgat gctccgagat
     3541 gaggtaggtg gtcgcgcggt taacttgctt cctagtgaaa ccgttcagga catctacggg
     3601 attgttgcta agaaagtcaa cgagattcta caagcagacg caatcaatgg gaccgataac
     3661 gaagtagtta ccgtgaccga tgagaacact ggtgaaatct ctgagaaagt caagctgggc
     3721 actaaggcac tggctggtca atggctggct tacggtgtta ctcgcagtgt gactaagcgt
     3781 tcagtcatga cgctggctta cgggtccaaa gagttcggct tccgtcaaca agtgctggaa
     3841 gataccattc agccagctat tgattccggc aagggtctga tgttcactca gccgaatcag
     3901 gctgctggat acatggctaa gctgatttgg gaatctgtga gcgtgacggt ggtagctgcg
     3961 gttgaagcaa tgaactggct taagtctgct gctaagctgc tggctgctga ggtcaaagat
     4021 aagaagactg gagagattct tcgcaagcgt tgcgctgtgc attgggtaac tcctgatggt
     4081 ttccctgtgt ggcaggaata caagaagcct attcagacgc gcttgaacct gatgttcctc
     4141 ggtcagttcc gcttacagcc taccattaac accaacaaag atagcgagat tgatgcacac
     4201 aaacaggagt ctggtatcgc tcctaacttt gtacacagcc aagacggtag ccaccttcgt
     4261 aagactgtag tgtgggcaca cgagaagtac ggaatcgaat cttttgcact gattcacgac
     4321 tccttcggta ccattccggc tgacgctgcg aacctgttca aagcagtgcg cgaaactatg
     4381 gttgacacct atgagtcttg tgatgtactg gctgatttct acgaccagtt cgctgaccag
     4441 ttgcacgagt ctcaattgga caaaatgcca gcacttccgg ctaaaggtaa cttgaacctc
     4501 cgtgacatct tagagtcgga cttcgcgttc gcgtaactgc agtataagct tggctgtttt
     4561 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     4621 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     4681 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     4741 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     4801 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     4861 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     4921 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt
     4981 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5041 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5101 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5161 taaaagatgc tgaagatcag ttgggtgcag caaactatta actggcgaac tacttactct
     5221 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5281 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5341 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5401 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5461 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5521 tgatttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5581 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5641 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5701 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     5761 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5821 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     5881 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5941 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag
     6001 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6061 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6301 ctcgctctgc taatcctgtt accagtcagg catttgagaa gcacacggtc acactgcttc
     6361 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     6421 ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc
     6481 acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta
     6541 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     6601 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     6661 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     6721 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     6781 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     6841 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     6901 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     6961 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat
     7021 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     7081 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     7141 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     7201 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     7261 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
     7321 gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg
     7381 atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact
     7441 gatttagtgt atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc
     7501 cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc
     7561 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
     7621 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     7681 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     7741 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     7801 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     7861 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     7921 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     7981 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     8041 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     8101 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     8161 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     8221 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     8281 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     8341 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     8401 tttctaggct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     8461 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     8521 agaggatctg ctcatgtttg acagcttatc
//
