LOCUS       sequence-451534-        4776 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding ClbS from Dickeya dadantii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4776)
  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 4776)
  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_Dickeya
FEATURES             Location/Qualifiers
     source          1..4776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1101..1106,1107..1123,1124..1130)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1138..1154
                     /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     1143..1165
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    1171..1240
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             1291..1317
                     /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(1868..1885)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2038..2124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2216..2243
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2262..2353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2354..2422,2423..3214)
                     /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(2572..2591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3385..3973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3874..3893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4127..4144
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4159..4299
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4225..4244)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4385..4407
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4485..4562
                     /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             join(4563..4776,1..869)
                     /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(4582..4601)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
       61 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
      121 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
      181 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
      241 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
      301 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
      361 tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc
      421 cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa
      481 tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg
      541 gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg
      601 gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg
      661 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
      721 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc
      781 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
      841 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagcgcgaa
      901 ttgatctggt ttgacagctt atcatcgact gcacggtgca ccaatgcttc tggcgtcagg
      961 cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg
     1021 ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc
     1081 aaatattctg aaatgagctg ttgacaatta atcatccggc tcgtataatg tgtggaattg
     1141 tgagcggata acaatttcac acaggaaaca gcgccgctga gaaaaagcga agcggcactg
     1201 ctctttaaca atttatcaga caatctgtgt gggcactcga ccggaattat cgattaactt
     1261 tattattaaa aattaaagag gtatatatta atgtatcgat taaataagga ggaataaacc
     1321 atgggcgtgc cacaaacaaa atccgaacta cttttcgcta ttgataaaaa ttttacgaaa
     1381 ttaatcggtt acctcaatgc gattccgcca gagctcactg cggataaatc gctggatggc
     1441 catgcaaaag gcacggagat gagtgtttgt gagcttgttt cgtatttgct gggatggaac
     1501 actctggttg taaaatggat tagcggtgac gcaacagggc aacctgttga tttcccggaa
     1561 accggttaca agtggaatca gctcgggctt ctggcgcaga aattttactt tgattacaaa
     1621 gaattaaatt accagacgtt aatcaataac cttcagtcgg taaaagacga aatagtgagg
     1681 ctgattgatg agcgcactga tgaagatctg tatggcaagc catggtatgg caaatggaca
     1741 atgggacgta tgatctcatt gaacacgtca tcaccctatg ccaatgctaa tggaagatta
     1801 aggcagtggg caaagaacaa tcacctgcgt ttaaaatgag ttttggcgga tgagagaaga
     1861 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     1921 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     1981 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     2041 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2101 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     2161 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     2221 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     2281 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2341 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2401 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2461 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2521 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2581 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     2641 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2701 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2761 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2821 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2881 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2941 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3001 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3061 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3121 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3181 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3241 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3301 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     3541 gtagccgtag ttaggccacc acttcaagaa ctctttagca ccgcctacat acctcgctct
     3601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4021 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4081 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4141 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4201 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4261 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4321 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4381 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4441 cagatcaatt cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc atcgaatggt
     4501 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     4561 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     4621 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     4681 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     4741 agtcgttgct gattggcgtt gccacctcca gtctgg
//
