LOCUS       sequence-451553-        4770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding ClbS from Samsonia erythrinae DSM 16730.
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_Samsonia
FEATURES             Location/Qualifiers
     source          1..4770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      117..203
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      295..322
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        341..432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(433..501,502..1293)
                     /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     complement(651..670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1464..2052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1953..1972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2206..2223
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2238..2378
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2304..2323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2464..2486
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2564..2641
                     /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."
     CDS             2642..3724
                     /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     complement(2661..2680)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3956..3961,3962..3978,3979..3985)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3993..4009
                     /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)."
     primer_bind     3998..4020
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    4026..4095
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             4146..4172
                     /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"
     primer_bind     complement(4717..4734)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
       61 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
      121 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      181 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
      241 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
      301 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata
      361 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      421 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      481 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      541 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      601 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      661 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      721 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      781 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
      841 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
      901 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
      961 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1021 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1081 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1141 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1201 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1261 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1321 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1381 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1441 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1501 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1561 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1621 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1681 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1741 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1801 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1861 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1921 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1981 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2041 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2101 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2161 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2221 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2281 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2341 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2401 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2461 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2521 agatcaattc gcgcgcgaag gcgaagcggc atgcatttac gttgacacca tcgaatggtg
     2581 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
     2641 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     2701 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     2761 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     2821 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     2881 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     2941 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     3001 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     3061 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     3121 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     3181 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     3241 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     3301 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     3361 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     3421 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     3481 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     3541 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     3601 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     3661 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3721 gtgagcgcaa cgcaattaat gtaagttagc gcgaattgat ctggtttgac agcttatcat
     3781 cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc
     3841 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
     3901 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
     3961 aattaatcat ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
     4021 aaacagcgcc gctgagaaaa agcgaagcgg cactgctctt taacaattta tcagacaatc
     4081 tgtgtgggca ctcgaccgga attatcgatt aactttatta ttaaaaatta aagaggtata
     4141 tattaatgta tcgattaaat aaggaggaat aaaccatggc ggtgccagaa tcaaaagatg
     4201 aactcattaa agcgatcaat agtcactttg cgttattgca gaaaaagctg ggggcagtcc
     4261 tgcctgagca gtcctttgaa ccgacaatgg aagggcatgc aaaagggagc gtgatgagcg
     4321 tggcccaatt ggtgtcttat cttattgggt ggggagaatt ggtcttgcac tggcacgaac
     4381 aagaagcgaa aggggaaacg ataaactttc ctgaagaggg ctataaatgg aatgaattgg
     4441 ggcgcctggc gcagaagttt tatcgcgact atgaagacat aaaggattat gaaatcttat
     4501 tgttgcgatt aaaggaaaac aaacagcagc ttttgacgct gattgagcgc tttagtaacg
     4561 aggaacttta cggtagtcct tggtatggca aatggacgcg ggggcgaatg attcaattta
     4621 atacctcctc gccttataaa aatgcttcag ggcgattaaa taaattgaaa aaacaggctc
     4681 cgatatgagt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
     4741 cagaagcggt ctgataaaac agaatttgcc
//
