LOCUS       sequence-484679-        7563 bp    DNA     circular SYN 26-AUG-2025
DEFINITION  AraC compatible Mutagenesis Circuit (drift plasmid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7563)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 7563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pLlacO1-DP6
FEATURES             Location/Qualifiers
     source          1..7563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             115..774
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAKCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(181..200)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(790..821)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(848..1474)
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             complement(1504..1758)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     protein_bind    4574..4590
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     terminator      4834..4920
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5012..5039
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 acggcctatt tggcttggca cgtaagaggt tccaactttc accataatga aataagatca
       61 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag
      121 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
      181 gaggcatttc agtcagttgc taaatgtacc tataaccaga ccgttcagct ggatattacg
      241 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
      301 cttgcccgcc tgatgaatgc tcatccggag ttccgtatgg caatgaaaga cggtgagctg
      361 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
      421 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
      481 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
      541 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
      601 atggacaact tcttcgcccc cgttttcact atgggcaaat attatacgca aggcgacaag
      661 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
      721 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcca
      781 tgggcatgta gtcaaaagcc tccggtcgga ggcttttgac ttggctgagg agtgccgtta
      841 attaagttta aaccgccgga gatttggtgg tgtgcaggat tttaacctgg atcatgatag
      901 acagttcaga acgacgtttt tccgcacgtt tcagggtttt ttccagccaa cggttttcgt
      961 tcagctggtt gtgagaagac tggatgaaga ttttacggca gcactggtag tgttcagaaa
     1021 ccataacgtt cagaccaaca ccgttgtcac gcaggttcca cagaccgatc tggttacgcg
     1081 cgtttttttc gtagtacagt ttgcacgccc agattttcag ggtgtgaccg ttaccacgca
     1141 gttcctggtt gtaccattcc aggatttttt ccgcgcagtc cgcgcacgga gaccaagaag
     1201 agtaccagtt gatggtgaac tgacccgggt tgtcacgcag gtattcttca actttacgga
     1261 tagagaagat ttccgcgtgg ataccacgtt cggtaccaga ctgcggtttg ttaaccgcgt
     1321 aaccccagaa gcacgcacga cgttcaccac gacgtttcag ttcgaacaga acgtagcaac
     1381 ggtgagaaac agattttttg ttgttgaaga actgtttttt gaaggtgtag atgtccagtt
     1441 tttcgtggat acgaacgtat tccgcgtcgg tcatcttcag attccggttt acttaatctc
     1501 gacttataac attttaattt tattttctcc attactgtct tgaattacta aagcccaagg
     1561 tttatattct ggagcatctg aagttaatag cattacattt tcatctgtac tttcatcata
     1621 agcagtatga actaaaatat cactttctgg tttattccca attacttcct ctacttcttc
     1681 tggtaacatt agaattgatt cttgaatcac tagttgtttt cctgtttctt tttcaatgat
     1741 gtcagataaa tttgtcatgt tgaaattcct cctaattttt gtattattag ctcatcgctt
     1801 cgagaaccac accgtcttgt tccatctggt cgagacggga gagcaatttg cgggtgattt
     1861 gctcgagctg atctttttct gttgtgctga gcgcggacca gagttgatgc aggcagttat
     1921 gctgcggtgg taaaacctcg cgcaaaaact cgtgaccttt ttccgttaat tgcagatgca
     1981 ggcagcggcg atcgttatcg ctttcacgac gttcgatcca accgcgtttt tccagttcat
     2041 cggcaatacg cgtcgcgttg gtacgggatg atccaagagc acaacttaat tcagaaggct
     2101 gaatactgtg gttttcctga gactccagcg taatcaacgc cataaacaac gtctcgttaa
     2161 ttccctgagc cttcagcatt ttattgcggt tctccagcag cttgctttgc atgtgcatgc
     2221 aaagacgagt cagaaggatc tcctgataag gaaaatcttc gtggcggctg gcgcgaaatt
     2281 ttagcatttg ttcaatgggc gtaaacgaac tatccatttg ggtatgacct cattaatttc
     2341 agccgttaga tagttccgca aaccttctca atcaattccg ccgggaattg catcgactgc
     2401 atgatgtgtt cgatcatgct gcatttacgg ccggtgttgg tgttggtgat cacccaatac
     2461 ggcgtgcctg gcacatgttt cggcttggtc tgattaccat ttttcagcag cgtttgttca
     2521 tctgccgcaa agtaaacgcg tgtacgaccg tgcaacgatt ccgttgcttc ggcaaacgcc
     2581 tgggcgtcaa gagaatatag tgtagacaac agcagcataa agcgattgac cgctcgcttt
     2641 tgctctgcgt attcatccga aagcagaagt tcacgcattg cgcgaacctt gtctttaatc
     2701 gttttgaccg gcttcgcttc gacgatagca ggtgacgcaa cgcgaacctc tttcgtcacc
     2761 ggagcagcag gctgtgatgc ggcggaaaat ttcaacatac gccgtaaaat gtcggatgcg
     2821 ctctcgccga tatgcttagt gtggctggca atatagctgt agagttcatc atcaacttca
     2881 atcgttttca tcttaatcca gtgcagtgtc ttagctgaat tattttttcg cgggtgaaac
     2941 gactcctggt ttgtacaaag ccagcagttc gtccaccttt ttacgtgtgc cgccgttgct
     3001 gcttatactg cgtcgaactt tgacgacatg caattttgcg cgctgatacc actcacgcgt
     3061 taacatcgta tcgtgattgg agatcagcac tggaatatgg cgctcaacca gaccttcggc
     3121 gatctccgcc agatgcgctt gttgttcaag cgtaaaactg tttgtgtgat acgccgtaaa
     3181 gttggcggtc gcagacagcg gtgcataagg cggatcgcaa tagacgacgg atgcatcatc
     3241 tgcgcgcgcc atgctatcgg cgtaagactc acaatagaaa aaggcattct gcgctttttc
     3301 agcgaagtga tacaactctg cttccgggaa atagggtttt ttgtagcggc cgaacggcac
     3361 gttaaactca ccgcgcagat tgtaacgaca caggccgttg taaccgtagc ggttcaaata
     3421 taaaaacagt accgcccgac ggaacggatc ctggcttttg ttgaactctt cgcggaactg
     3481 atagtaaacc tcggcgcaat ttgtttcggg aacaaacagc tcgcgtgcgg cctgtacgta
     3541 ctcatcagta cgcatcttca caatgttata gagactgatc aggtcgctat tgatatcggc
     3601 aaggatataa cgagaaaagt cggtgttgag aaacaccgac ccggcaccta caaaaggctc
     3661 aaccagacat tcgcccttgg gcaaatgccg tttaatatca tcaagcaggg gatacttgcc
     3721 ccctgccact tcaaaaaagc gcgatttttc ttcatgggtt atgtccggtt tactgatatt
     3781 aaattatgct cgccagaggc aacttccgcc tttcttctgc accagatcga gacgggcttc
     3841 atgagctgca atctcttcat ctgtcgcaaa aacaacgcgt aacttacttg cctgacgtac
     3901 aatgcgctga attgttgctt caccttgttg ctgttgtgtc tctccttcca tcgcaaaagc
     3961 catcgacgtt tgaccaccgg tcatcgccag ataaacttcc gcaaggatct gggcatcgag
     4021 taatgccccg tgcagcgttc gtttactgtt atctatttcg tagcgagcac ataacgcatc
     4081 gaggctgttg cgcttaccgg gaaacatttt cctcgccacc gcaaggctat cggtgacctt
     4141 acagaaagta ttggtcttcg gaatatcgcg cttaagcaac gaaaactcgt agtccataaa
     4201 gccgatatcg aacgctgcgt tatggatcac caactccgcg ccgcgaatat agtccatgaa
     4261 ctcatcggct acttcggcaa acgtgggctt atcgagcaaa aattcatcgg caataccatg
     4321 tacgccaaag gcttccggat ccaccagccg atcgggtttg agataaacat ggaagttatt
     4381 gcccgtcagg cgacggttca ccacttcaac ggcaccaatc tcaatgatct tgtggccttc
     4441 atagtgcgca ccaatctggt tcataccggt ggttgcggta gcgagaacga tctggcgtgt
     4501 aattgcagtg ctcatctagt aggtttcctg tgtgagtctt cggtcagtgc gtcctgctga
     4561 tgtgctcagt atcttgttat ccgctcacaa tgtcaatgtt atccgctcac atttataggg
     4621 tccctaagtc tggctgtttt ggcggatgag agaagatttt cagcctgata cagattgaat
     4681 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     4741 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     4801 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     4861 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     4921 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     4981 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     5041 gcgtttctac aaactcgctg aggacattgg ctccgtgatg aaattcgcca cttgttagtg
     5101 taattcgcta actcatcctg gcatgttgct gttgattctt caatcagatc tttataaatc
     5161 aaaaagataa aaaattggca cgcaaattgt attaacagtt catccctatc agtgatagat
     5221 agcaggacaa tcctgaacgc agaaatcaag aggacaacat tacctgcagg tgcagtaaag
     5281 gaaaaaaaaa atgaaaaaat tattattcgc aattccttta gttgttcctt tctattctca
     5341 ctccgctgaa actgttgaaa gttgtttagc aaaaccccat acagaaaatt catttactaa
     5401 cgtctggaaa gacgacaaaa ctttagatcg ttacgctaac tatgagggct gtctgtggaa
     5461 tgctacaggc gttgtagttt gtactggtga cgaaactcag tgttacggta catgggttcc
     5521 tattgggctt gctatccctg aaaatgaggg tggtggctct gagggtggcg gttctgaggg
     5581 tggcggctct gagggtggcg gtactaaacc tcctgagtac ggtgatacac ctattccggg
     5641 ctatacttat atcaaccctc tcgacggcac ttatccgcct ggtactgagc aaaaccccgc
     5701 taatcctaat ccttctcttg aggagtctca gcctcttaat actttcatgt ttcagaataa
     5761 taggttccga aataggcagg gggcattaac tgtttatacg ggcactgtta ctcaaggcac
     5821 tgaccccgtt aaaacttatt accagtacac tcctgtatca tcaaaagcca tgtatgacgc
     5881 ttactggaac ggtaaattca gagactgcgc tttccattct ggctttaatg aggatccatt
     5941 cgtttgtgaa tatcaaggcc aatcgtctga cctgcctcaa cctcctgtca atgctggcgg
     6001 cggctctggt ggtggttctg gtggcggctc tgagggtggt ggctctgagg gtggcggttc
     6061 tgagggtggc ggctctgagg gaggcggttc cggtggtggc tctggttccg gtgattttga
     6121 ttatgaaaag atggcaaacg ctaataaggg ggctatgacc gaaaatgccg atgaaaacgc
     6181 gctacagtct gacgctaaag gcaaacttga ttctgtcgct actgattacg gtgctgctat
     6241 cgatggtttc attggtgacg tttccggcct tgctaatggt aatggtgcta ctggtgattt
     6301 tgctggctct aattcccaaa tggctcaagt cggtgacggt gataattcac ctttaatgaa
     6361 taatttccgt caatatttac cttccctccc tcaatcggtt gaatgtcgcc cttttgtctt
     6421 tggcgctggt aaaccttacg agttcagtat cgactgcgat aagatcaacc tgttccgcgg
     6481 tgtctttgcg tttcttttat atgttgccac ctttatgtat gtattttcta cgtttgctaa
     6541 catactgcgt aataaggagt cttaaagggt ccctaagtct cctcagcaaa acgaaaggcc
     6601 cagtctttcg actgagcctt tcgttttatt tgaccggatg tcctcttcct cctgggctga
     6661 cttcaggtgc tacatttgaa gagataaact gcactgaaat ctagagcggt tcagtagaaa
     6721 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatcttttgc cctgtaaacg
     6781 aaaaaaccac ctggggaggt ggtttgatcg aaggttaagc agttcaccaa ctgacttaac
     6841 cttcgaaaaa accacctggg gaggtggttt gatcgaaggt taagtcagtt ggggaactgc
     6901 ttaaccgtgg taactggctt tcgcagagca cagcaaccaa atctgtcctt ccagtgtagc
     6961 cggactttgg cgcacacttc aagagcaacc gcgtgtttag ctaaacaaat cctctgcgaa
     7021 ctcccagtta ccaatggctg ctgccagtgg cgttttaccg tgcttttccg ggttggactc
     7081 aagtgaacag ttaccggata aggcgcagca gtcgggctga acggggagtt cttgcttaca
     7141 gcccagcttg gagcgaacga cctacaccga gccgagatac cagtgtgtga gctatgagaa
     7201 agcgccacac ttcccgtaag ggagaaaggc ggaacaggta tccggtaaac ggcagggtcg
     7261 gaacaggaga gcgcaagagg gagcgacccg ccggaaacgg tggggatctt taagtcctgt
     7321 cgggtttcgc ccgtactgtc agattcatgg ttgagcctca cggctcccac agatgcaccg
     7381 gaaaagcgtc tgtttatgtg aactctggca ggagggcgga gcctatggaa aaacgccacc
     7441 ggcgcggccc tgctgttttg cctcacatgt tagtcccctg cttatccacg gaatctgtgg
     7501 gtaactttgt atgtgtccgc agcgcccgcc gcagtctcac gcccggagcg tagcgaccga
     7561 gtg
//
