LOCUS       sequence-451540-        4770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding ClbS from Snodgrassella alvi wkB2.
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_Snod
FEATURES             Location/Qualifiers
     source          1..4770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..65
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(85..162)
                     /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(240..262)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    348..488
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     403..422
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(503..520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(674..1262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(754..773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1433..2224,2225..2293))
                     /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     2056..2075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2294..2385)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2404..2431)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2523..2609)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3324..3350)
                     /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    3401..3470
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     primer_bind     complement(3476..3498)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3487..3503
                     /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(3511..3517,3518..3534,3535..3540))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(join(3772..4770,1..84))
                     /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 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
       61 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
      121 tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcaacgtaaa tgcatgccgc
      181 ttcgccttcg cgcgcgaatt gatctgctgc ctcgcgcgtt tcggtgatga cggtgaaaac
      241 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
      301 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
      361 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
      421 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
      481 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1081 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     1141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1381 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1501 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1621 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1861 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1921 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2041 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2101 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2281 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt ttgtagaaac
     2401 gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc agtttatggc
     2461 gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc cgctcccggc
     2521 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     2581 tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg
     2641 agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg
     2701 tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc ttctgcgttc
     2761 tgatttaatc cgtatcaggc tgaaaatctt ctctcatccg ccaaaactta ttctttgatc
     2821 tgtttcaaca atttattcaa tcgtcctgtg gcatttttat aaggggagga ggtattaaac
     2881 tgaatcatgc gtccgcgtgt gtatttttca tgccagggtg agccatacag ctcttcatta
     2941 ctaaagctat caataagtga aatcagattc tttttgttgg tctccagaag cgagagtaac
     3001 tgtttgaaat cggtaatgtc ctgataatca cgataaaatt tctgtgccag taatcccagt
     3061 tgattccatt tatatcctgc ttccggaaag gcaattttct gccctgactg ttcctgttta
     3121 tgccatgaca gcaccagttc tccccagcca attaaataag acacaagctg tgcagcactc
     3181 atcattgtac ctttagcatg gccttccatc agaggcagat aggctttctg ttcaggtaca
     3241 gcagccagct ttttggtcag tagtgtatag tttttattaa ttgcttcaat tagttcctgt
     3301 tttgattctg gtattgccat ggtttattcc tccttattta atcgatacat taatatatac
     3361 ctctttaatt tttaataata aagttaatcg ataattccgg tcgagtgccc acacagattg
     3421 tctgataaat tgttaaagag cagtgccgct tcgctttttc tcagcggcgc tgtttcctgt
     3481 gtgaaattgt tatccgctca caattccaca cattatacga gccggatgat taattgtcaa
     3541 cagctcattt cagaatattt gccagaaccg ttatgatgtc ggcgcaaaaa acattatcca
     3601 gaacgggagt gcgccttgag cgacacgaat tatgcagtga tttacgacct gcacagccat
     3661 accacagctt ccgatggctg cctgacgcca gaagcattgg tgcaccgtgc agtcgatgat
     3721 aagctgtcaa accagatcaa ttcgcgctaa cttacattaa ttgcgttgcg ctcactgccc
     3781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3841 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     3901 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     3961 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     4021 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     4081 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     4141 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     4201 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     4261 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     4321 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     4381 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     4441 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     4501 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     4561 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     4621 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     4681 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     4741 cactttttcc cgcgttttcg cagaaacgtg
//
