LOCUS       sequence-451541-        4773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding ClbS from Gibbsiella quercinecans DSM 25889.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4773)
  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 4773)
  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_Gibsiella
FEATURES             Location/Qualifiers
     source          1..4773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     384..403
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(423..500)
                     /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(578..600)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    686..826
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(841..858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1012..1600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1092..1111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1771..2562,2563..2631))
                     /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     2394..2413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2632..2723)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(2742..2769)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2861..2947)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3100..3117
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(3665..3691)
                     /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    3742..3811
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     primer_bind     complement(3817..3839)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3828..3844
                     /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(3852..3858,3859..3875,3876..3881))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(join(4113..4773,1..422))
                     /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 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
       61 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
      121 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
      181 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
      241 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
      301 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
      361 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
      421 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
      481 ttgcaccatt cgatggtgtc aacgtaaatg catgccgctt cgccttcgcg cgcgaattga
      541 tctgctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      601 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      661 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
      721 tgtatactgg cttaactatg ctgcatcaga gcagattgta ctgagagtgc accatatgcg
      781 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
      841 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      901 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      961 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1021 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1081 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1141 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1201 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1261 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1321 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1381 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1441 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1501 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1561 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1621 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1681 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1741 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1801 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1861 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1921 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1981 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2041 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2101 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2161 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2221 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2281 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2341 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac
     2401 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2461 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2521 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2581 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2641 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2701 atgtatttag aaaaataaac aaaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     2761 tggccttctg cttaatttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     2821 cgggccgttg cttcgcaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     2881 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     2941 ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact accatcggcg
     3001 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg
     3061 ccaggcaaat tctgttttat cagaccgctt ctgcgttctg atttaatctg tatcaggctg
     3121 aaaatcttct ctcatccgcc aaaacttacg ggacacagcc tacttttcgc aatgcgttta
     3181 gcctgccgca ggcgttctta tacggtgacg ccgtgttgaa ctgaatcatc cggccacgcg
     3241 tccatttgcc gtaccacggc tggccgtaga gcgcctcatc gctaaagccg tcaatcaacg
     3301 cgataatctg ccgtttatta tcggcaagcc gcatcagcag ggcgccgtag tccgtaatct
     3361 gcgcataatc acggtaaaac ttttgcgcca gcaaccccaa ttggttccac ttaaaaccgt
     3421 cttcggggaa ggcgatttca aggccctgtc tttctttcgc atgccaggac aaaaccagtt
     3481 ccccccagcc aatcagatag gccaccagtt gggcaacgct cattacgctg cccttggcat
     3541 gccccggcat cagcggctca aacgcctttt ctgccggtat gtcgtccagt tttttcatca
     3601 gcagcgcgaa attgctgttc accgctttga tcaatgcgcc ttttgattcc ggcacggcca
     3661 tggtttattc ctccttattt aatcgataca ttaatatata cctctttaat ttttaataat
     3721 aaagttaatc gataattccg gtcgagtgcc cacacagatt gtctgataaa ttgttaaaga
     3781 gcagtgccgc ttcgcttttt ctcagcggcg ctgtttcctg tgtgaaattg ttatccgctc
     3841 acaattccac acattatacg agccggatga ttaattgtca acagctcatt tcagaatatt
     3901 tgccagaacc gttatgatgt cggcgcaaaa aacattatcc agaacgggag tgcgccttga
     3961 gcgacacgaa ttatgcagtg atttacgacc tgcacagcca taccacagct tccgatggct
     4021 gcctgacgcc agaagcattg gtgcaccgtg cagtcgatga taagctgtca aaccagatca
     4081 attcgcgcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4141 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4201 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     4261 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     4321 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     4381 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     4441 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     4501 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     4561 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     4621 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     4681 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     4741 tcagagacat caagaaataa cgccggaaca tta
//
