LOCUS       sequence-451554-        4770 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding ClbS from Frischella perrara.
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_Frischella
FEATURES             Location/Qualifiers
     source          1..4770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      36..63
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        82..173
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(174..242,243..1034)
                     /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(392..411)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1205..1793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1694..1713
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1947..1964
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1979..2119
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2045..2064)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2205..2227
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2305..2382
                     /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             2383..3465
                     /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(2402..2421)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3697..3702,3703..3719,3720..3726)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3734..3750
                     /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     3739..3761
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    3767..3836
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             3887..3913
                     /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(4458..4475)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4628..4714
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
       61 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc
      121 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      181 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      241 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      301 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      361 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
      421 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      481 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      541 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      601 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      661 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
      721 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      781 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      841 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      901 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      961 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1021 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1081 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1141 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1201 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1261 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1321 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     1381 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1441 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1501 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1561 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1621 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1681 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1741 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1801 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1861 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1921 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     1981 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     2041 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     2101 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gcygacgcgc cctgacgggc
     2161 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2221 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
     2281 ggcgaagcgg catgcattta cgttgacacc atcgaatggt gcaaaacctt tcgcggtatg
     2341 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     2401 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     2461 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     2521 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     2581 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     2641 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     2701 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     2761 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     2821 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     2881 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     2941 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     3001 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     3061 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     3121 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     3181 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     3241 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     3301 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     3361 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3421 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3481 tgtaagttag cgcgaattga tctggtttga cagcttatca tcgactgcac ggtgcaccaa
     3541 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
     3601 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
     3661 tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca tccggctcgt
     3721 ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcgc cgctgagaaa
     3781 aagcgaagcg gcactgctct ttaacaattt atcagacaat ctgtgtgggc actcgaccgg
     3841 aattatcgat taactttatt attaaaaatt aaagaggtat atattaatgt atcgattaaa
     3901 taaggaggaa taaaccatgg cagtacctga atctaaaact gagttaatag aagcgattaa
     3961 taaaaattat gcttcgttag tcaaaaagct atcagctgta ccagcagaaa aggctttcca
     4021 accattaatg gaggggcatg ccaaaggcac gactatgagt gcagcacaac ttgtttctta
     4081 tttaataggt tggggcgaat tagtcttatc ttggcatgca caagaagcac atgggcatag
     4141 catcgttttt cctgaagtgg gctttaaatg gaatgaatta ggaaaactag cacagaaatt
     4201 ttatcaagac tatcaaaaca tcactgatta caaggagcta ttacaacggt tagaaaaaaa
     4261 taaaaatgat cttattaaat tgattgaagg ttttacgaat aatgatttat atggttcgag
     4321 ttggtgtgaa aaatacactc gaggacgaat gattcagcta aatacgtcat caccgtatag
     4381 aaatgcaagt ggtcgcatcg cgacattcct caaaactatc ggcaattaag ttttggcgga
     4441 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     4501 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     4561 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     4621 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     4681 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     4741 gaagcaacgg cccggagggt ggcgggcagg
//
