LOCUS       sequence-128764-        5621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Untargetted Bacterial Mutagenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  AUTHORS   Badran AH, Liu DR
  TITLE     Development of potent in vivo mutagenesis plasmids with broad 
            mutational spectra.
  JOURNAL   Nat Commun. 2015 Oct 7;6:8425. doi: 10.1038/ncomms9425.
  PUBMED    26443021
REFERENCE   2  (bases 1 to 5621)
  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: "2015-10-7"; volume: "6:8425. doi"; pages: " 
            10.1038/ncomms9425"
COMMENT     Sequence Label: MP-H2
FEATURES             Location/Qualifiers
     source          1..5621
                     /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"
     terminator      4801..4832
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4848..5507)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5422..5441
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
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 ccgctatgag cactgcaatt acacgccaga tcgttctcgc taccgcaacc accggtatga
     2221 accagattgg tgcgcactat gaaggccaca agatcattga gattggtgcc gttgaagtgg
     2281 tgaaccgtcg cctgacgggc aataacttcc atgtttatct caaacccgat cggctggtgg
     2341 atccggaagc ctttggcgta catggtattg ccgatgaatt tttgctcgat aagcccacgt
     2401 ttgccgaagt agccgatgag ttcatggact atattcgcgg cgcggagttg gtgatccata
     2461 acgcagcgtt cgatatcggc tttatggact acgagttttc gttgcttaag cgcgatattc
     2521 cgaagaccaa tactttctgt aaggtcaccg atagccttgc ggtggcgagg aaaatgtttc
     2581 ccggtaagcg caacagcctc gatgcgttat gtgctcgcta cgaaatagat aacagtaaac
     2641 gaacgctgca cggggcatta ctcgatgccc agatccttgc ggaagtttat ctggcgatga
     2701 ccggtggtca aacgtcgatg gcttttgcga tggaaggaga gacacaacag caacaaggtg
     2761 aagcaacaat tcagcgcatt gtacgtcagg caagtaagtt acgcgttgtt tttgcgacag
     2821 atgaagagat tgcagctcat gaagcccgtc tcgatctggt gcagaagaaa ggcggaagtt
     2881 gcctctggcg agcataaatc gcacgctgcc aaactaagga cgattgatgc caattcaggt
     2941 cttaccgcca caactggcga accagattgc cgcaggtgag gtggtcgagc gacctgcgtc
     3001 ggtagtcaaa gaactagtgg aaaacagcct cgatgcaggt gcgacgcgta tcgatattga
     3061 tatcgaacgc ggtggggcga aacttatccg cattcgtgat aacggctgcg gtatcaaaaa
     3121 agatgagctg gcgctggcgc tggctcgtca tgccaccagt aaaatcgcct ctctggacga
     3181 tctcgaagcc attatcagcc tgggctttcg cgatgaggcg ctggcgagta tcagttcggt
     3241 ttcccgcctg acgctcactt cacgcaccgc agaacagcag gaagcctggc aggcctatgc
     3301 cgaagggcgc gatatgaacg tgacggtaaa accggcggcg catcctgtgg ggacgacgct
     3361 ggaggtgctg gatctgttct acaacacccc ggcgcggcgc aaattcctgc gcaccgagaa
     3421 aaccgaattt aaccacattg atgagatcat ccgccgcatt gcgctggcgc gtttcgacgt
     3481 cacgatcaac ctgtcgcata acggtaaaat tgtgcgtcag taccgcgcag tgccggaagg
     3541 cgggcaaaaa gaacggcgct taggcgcgat ttgcggcacc gcttttcttg aacaagcgct
     3601 ggcgattgaa tggcaacacg gcgatctcac gctacgcggc tgggtggccg atccaaatca
     3661 caccacgccc gcactggcag aaattcagta ttgctacgtg aacggtcgca tgatgcgcga
     3721 tcgcctgatc aatcacgcga tccgccaggc ctgcgaagac aaactggggg ccgatcagca
     3781 accggcattt gtgttgtatc tggagatcga cccacatcag gtggacgtca acgtgcaccc
     3841 cgccaaacac gaagtgcgtt tccatcagtc gcgtctggtg catgatttta tctatcaggg
     3901 cgtgctgagc gtgctacaac agcaactgga aacgccgcta ccgctggacg atgaacccca
     3961 acctgcaccg cgttccattc cggaaaaccg cgtggcggcg gggcgcaatc actttgcaga
     4021 accggcagct cgtgagccgg tagctccgcg ctacactcct gcgccagcat caggcagtcg
     4081 tccggctgcc ccctggccga atgcgcagcc aggctaccag aaacagcaag gtgaagtgta
     4141 tcgccagctt ttgcaaacgc ccgcgccgat gcaaaaatta aaagcgccgg aaccgcagga
     4201 acctgcactt gcggcgaaca gtcagagttt tggtcgggta ctgactatcg tccattccga
     4261 ctgtgcgttg ctggagcgcg acggcaacat ttcactttta tccttgccag tggcagaacg
     4321 ttggctgcgt caggcacaat tgacgccggg tgaagcgccc gtttgcgccc agccgctgct
     4381 gattccgttg cggctaaaag tttctgccga agaaaaatcg gcattagaaa aagcgcagtc
     4441 tgccctggcg gaattgggta ttgatttcca gtcagatgca cagcatgtga ccatcagggc
     4501 agtgccttta cccttacgcc aacaaaattt acaaatcttg attcctgaac tgataggcta
     4561 cctggcgaag cagtccgtat tcgaacctgg caatattgcg cagtggattg cacgaaattt
     4621 gatgagcgaa catgcgcagt ggtcaatggc acaggccata accctgctgg cggacgtgga
     4681 acggttatgt ccgcaacttg tgaaaacgcc gccgggtggt ctgttacaat ctgttgattt
     4741 acatccggcg ataaaagccc tgaaagatga gtgaacttaa ttaacggcac tcctcagcca
     4801 agtcaaaagc ctccgaccgg aggcttttga ctacatgccc atggcgttta cgccccgccc
     4861 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     4921 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     4981 tatttgccca tagtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     5041 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     5101 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     5161 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     5221 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     5281 gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     5341 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     5401 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     5461 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     5521 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     5581 tgaaagttgg aacctcttac gtgccaagcc aaataggccg t
//
