LOCUS       sequence-451557-        4764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding ClbS from Escherichia albertii CB9786.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4764)
  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 4764)
  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_Ealbertii
FEATURES             Location/Qualifiers
     source          1..4764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..58
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(597..623)
                     /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    674..743
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     primer_bind     complement(749..771)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    760..776
                     /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(784..790,791..807,808..813))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(1045..2127)
                     /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     2089..2108
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2128..2205)
                     /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(2283..2305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2391..2531
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2446..2465
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2546..2563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2717..3305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2797..2816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3476..4267,4268..4336))
                     /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     4099..4118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4337..4428)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(4447..4474)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4566..4652)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gccaggcaaa ttctgtttta tcagaccgct tctgcgttct gatttaatct gtatcaggct
       61 gaaaatcttc tctcatccgc caaaactcag aattgtttaa gtagggcgtt cagacgtgtg
      121 cttgcatttc tatagggtga tacggtatta aattggatca ttcgaccacg cgtccatttg
      181 ccataccagg gatgatggta taactcgtta tcgctgtagc gttccactag cgatttgagt
      241 tgctgatgat tggtttctaa catttcgcac aattgggacc atgatgtgac atgggaataa
      301 ttagcgtaaa atgtctgcgc tagtttccca agttcattcc acttatggtc tttagccgga
      361 aaatcaatgg ggattcctgc ttcttcttgt ctgtgccagg taagaacctg ctctccccaa
      421 cctattaaat agcttaccag gttagccgca ctcatcacag tgccgctggc atgtccttcc
      481 attgtgatga gaaaagcttt ttcttctgga atgcgagata acttttttgt aagtccattc
      541 catgattttt ccatttccag caacagtgca gctttgcttt ctggtacaga catggtttat
      601 tcctccttat ttaatcgata cattaatata tacctcttta atttttaata ataaagttaa
      661 tcgataattc cggtcgagtg cccacacaga ttgtctgata aattgttaaa gagcagtgcc
      721 gcttcgcttt ttctcagcgg cgctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
      781 acacattata cgagccggat gattaattgt caacagctca tttcagaata tttgccagaa
      841 ccgttatgat gtcggcgcaa aaaacattat ccagaacggg agtgcgcctt gagcgacacg
      901 aattatgcag tgatttacga cctgcacagc cataccacag cttccgatgg ctgcctgacg
      961 ccagaagcat tggtgcaccg tgcagtcgat gataagctgt caaaccagat caattcgcgc
     1021 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1081 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     1141 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     1201 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     1261 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     1321 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     1381 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     1441 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     1501 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     1561 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     1621 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     1681 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     1741 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     1801 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     1861 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     1921 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     1981 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     2041 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     2101 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     2161 ctatcatgcc ataccgcgaa aggttttgca ccattcgatg gtgtcaacgt aaatgcatgc
     2221 cgcttcgcct tcgcgcgcga attgatctgc tgcctcgcgc gtttcggtga tgacggtgaa
     2281 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2341 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     2401 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     2461 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2521 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2581 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2641 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2701 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2761 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2821 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2881 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2941 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3001 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3061 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3121 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3181 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3241 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3361 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3421 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3541 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3661 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3721 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3901 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4021 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4081 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4141 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4201 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4321 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4381 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaaag agtttgtaga
     4441 aacgcaaaaa ggccatccgt caggatggcc ttctgcttaa tttgatgcct ggcagtttat
     4501 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttcg caacgttcaa atccgctccc
     4561 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     4621 cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg
     4681 gggagacccc acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc
     4741 aggtgggacc accgcgctac tgcc
//
