LOCUS       sequence-451539-        4770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding ClbS from Mixta theicola.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4770)
  AUTHORS   Silpe JE, Wong JWH, Owen SV, Baym M, Balskus EP
  TITLE     The bacterial toxin colibactin triggers prophage induction.
  JOURNAL   Nature. 2022 Mar;603(7900):315-320. doi: 10.1038/s41586-022-04444-3.
            Epub 2022 Feb 23.
  PUBMED    35197633
REFERENCE   2  (bases 1 to 4770)
  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"; doi: 
            "10.1038/s41586-022-04444-3"; journalName: "Nature"; date: 
            "2022-03"; volume: "603"; issue: "7900"; pages: "315-320"
COMMENT     Sequence Label: pTrc-clbS_Mixta
FEATURES             Location/Qualifiers
     source          1..4770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     820..839
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(859..936)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(1014..1036)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1122..1262
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1177..1196
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1277..1294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1448..2036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1528..1547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2207..2998,2999..3067))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2830..2849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3068..3159)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(3178..3205)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3297..3383)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3536..3553
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(4098..4124)
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     misc_feature    4175..4244
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     primer_bind     complement(4250..4272)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4261..4277
                     /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)."
     promoter        complement(join(4285..4291,4292..4308,4309..4314))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(join(4546..4770,1..858))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
       61 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
      121 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
      181 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
      241 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
      301 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
      361 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
      421 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
      481 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
      541 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
      601 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
      661 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
      721 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
      781 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
      841 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
      901 cataccgcga aaggttttgc accattcgat ggtgtcaacg taaatgcatg ccgcttcgcc
      961 ttcgcgcgcg aattgatctg ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     1021 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     1081 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     1141 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     1201 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     1261 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     1321 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1381 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1441 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1501 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1561 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1621 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1681 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1741 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1801 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1861 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     1921 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1981 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     2401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2821 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3121 gcggatacat atttgaatgt atttagaaaa ataaacaaaa gagtttgtag aaacgcaaaa
     3181 aggccatccg tcaggatggc cttctgctta atttgatgcc tggcagttta tggcgggcgt
     3241 cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc cggcggattt
     3301 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     3361 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat ggggagaccc
     3421 cacactacca tcggcgctac ggcgtttcac ttctgagttc ggcatggggt caggtgggac
     3481 caccgcgcta ctgccgccag gcaaattctg ttttatcaga ccgcttctgc gttctgattt
     3541 aatctgtatc aggctgaaaa tcttctctca tccgccaaaa cttacgcagc agtttgtttt
     3601 ttcaatttgt ttaatcgtcc tgcggcattt ctatagggcg aggcggtatt aaattgaatc
     3661 attcgtcctc gcgtccattt gccataccag ggcgtaccgt aaagttcatc gttgctaaat
     3721 tgctcagtca gcatcaagag ccgctgtttg ttctccctta accgcgccaa caaggtttca
     3781 taattgttga tgtcttcata gtcacggtag aatttctgcg ccagctgtcc cagctcattc
     3841 catttatagc cctctgcagg aaacacgatg gtttcccctt tcgcctcttg gttatgccag
     3901 tgcaacacca attcgcccca gccaatcagc caggagacca gctgcgccac gctcatcgtg
     3961 ctccctttgg catgcccttc cataagcggc tcaaaggcaa gcgctggcgc aatagcctcc
     4021 agctttttat tcagcaacgc gaaattacta ttgatcgctt taattaattc agcttttgat
     4081 cccggtaccg ccatggttta ttcctcctta tttaatcgat acattaatat atacctcttt
     4141 aatttttaat aataaagtta atcgataatt ccggtcgagt gcccacacag attgtctgat
     4201 aaattgttaa agagcagtgc cgcttcgctt tttctcagcg gcgctgtttc ctgtgtgaaa
     4261 ttgttatccg ctcacaattc cacacattat acgagccgga tgattaattg tcaacagctc
     4321 atttcagaat atttgccaga accgttatga tgtcggcgca aaaaacatta tccagaacgg
     4381 gagtgcgcct tgagcgacac gaattatgca gtgatttacg acctgcacag ccataccaca
     4441 gcttccgatg gctgcctgac gccagaagca ttggtgcacc gtgcagtcga tgataagctg
     4501 tcaaaccaga tcaattcgcg ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     4561 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4621 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4681 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4741 cagcaggcga aaatcctgtt tgatggtggt
//
