LOCUS       sequence-451533-        4770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding wild type ClbS from E. coli CFT073.
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
FEATURES             Location/Qualifiers
     source          1..4770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(326..352)
                     /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    403..472
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     primer_bind     complement(478..500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    489..505
                     /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(513..519,520..536,537..542))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(774..1856)
                     /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"
     primer_bind     1818..1837
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1857..1934)
                     /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(2012..2034)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2120..2260
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2175..2194
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2275..2292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2446..3034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2526..2545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3205..3996,3997..4065))
                     /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     3828..3847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4066..4157)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4176..4203)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4295..4381)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4534..4551
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 aaacttcgta ctctgtaata tcctcatagt cacggtagaa tttctgtgct aaacgcccca
       61 attcattcca tttaaatcct tcctcaggaa aaataatagt ttttcctttt gcctcttggt
      121 catgccagtg taacaccagc tctccccagc caatcagata ggaaaccaga ttcgctacgc
      181 taatcgtagt ccccttcgcg tgcccttcca atagaggttc aaaggcgagt tggggcgtaa
      241 tagattctag cttcttattt aataaagaaa aattactatt aatagcttta attaactctt
      301 cttttgatga tggaacagcc atggtttatt cctccttatt taatcgatac attaatatat
      361 acctctttaa tttttaataa taaagttaat cgataattcc ggtcgagtgc ccacacagat
      421 tgtctgataa attgttaaag agcagtgccg cttcgctttt tctcagcggc gctgtttcct
      481 gtgtgaaatt gttatccgct cacaattcca cacattatac gagccggatg attaattgtc
      541 aacagctcat ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa aaacattatc
      601 cagaacggga gtgcgccttg agcgacacga attatgcagt gatttacgac ctgcacagcc
      661 ataccacagc ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt gcagtcgatg
      721 ataagctgtc aaaccagatc aattcgcgct aacttacatt aattgcgttg cgctcactgc
      781 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      841 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
      901 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
      961 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     1021 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     1081 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     1141 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     1201 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     1261 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     1321 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     1381 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     1441 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     1501 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     1561 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     1621 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     1681 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     1741 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     1801 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     1861 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac
     1921 cattcgatgg tgtcaacgta aatgcatgcc gcttcgcctt cgcgcgcgaa ttgatctgct
     1981 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     2041 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     2101 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     2161 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     2221 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     2281 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2341 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2401 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2461 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2521 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2581 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2641 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2701 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2761 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2821 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2881 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2941 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3001 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3061 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3121 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3181 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3241 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3301 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3361 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3421 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3481 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3541 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3601 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3661 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3721 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3781 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     3841 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3901 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3961 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4021 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4081 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4141 ttagaaaaat aaacaaaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     4201 tctgcttaat ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     4261 gttgcttcgc aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     4321 ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt
     4381 gatgcctggc agttccctac tctcgcatgg ggagacccca cactaccatc ggcgctacgg
     4441 cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact gccgccaggc
     4501 aaattctgtt ttatcagacc gcttctgcgt tctgatttaa tctgtatcag gctgaaaatc
     4561 ttctctcatc cgccaaaacc tattctgcaa gacatttctg cagtttattt aacctccccg
     4621 aagcattttt ataaggcgag gcggtattaa attgaatcat acgacctcgg gtccatttat
     4681 tataccaagg tttaccgtaa agctcgtcgt tactgaatcg ttcaatcaaa gccacgagtt
     4741 gctgcttatt ttcctttaac cgtgccaata
//
