LOCUS       sequence-451555-        4797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding ClbS from Bifidobacterium longum subsp. infantis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4797)
  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 4797)
  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_Bifido
FEATURES             Location/Qualifiers
     source          1..4797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(234..251)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      404..490
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      582..609
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        628..719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(720..788,789..1580)
                     /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(938..957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1751..2339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2240..2259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2493..2510
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2525..2665
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2591..2610)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2751..2773
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        2851..2928
                     /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             2929..4011
                     /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(2948..2967)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4243..4248,4249..4265,4266..4272)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4280..4296
                     /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     4285..4307
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    4313..4382
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             4433..4459
                     /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"
ORIGIN
        1 gttgaccgac aacgtcgacg ccatatacgc catgatcgat gcgatgagcg aggacgagct
       61 gttcaagccg catatgagga agtgggccga cgacgccacc aaaacggcgg tatgggaagt
      121 gtacaggttc attcatgtga acaccgtcgc cccgttcgga tcgttcagga cgaagatccg
      181 caagtggaag cggatggccc tataggtttt ggcggatgag agaagatttt cagcctgata
      241 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
      301 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
      361 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
      421 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
      481 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
      541 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
      601 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
      661 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      721 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
      781 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
      841 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
      901 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      961 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1021 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1081 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1141 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1201 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1261 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1321 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1381 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     1441 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     1501 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     1561 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1621 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1681 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1741 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1861 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     1921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2341 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2401 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     2461 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     2521 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     2581 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     2641 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     2701 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2761 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
     2821 cgcgaaggcg aagcggcatg catttacgtt gacaccatcg aatggtgcaa aacctttcgc
     2881 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     2941 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     3001 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     3061 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     3121 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     3181 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     3241 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     3301 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     3361 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     3421 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     3481 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     3541 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     3601 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     3661 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     3721 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     3781 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     3841 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     3901 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3961 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     4021 aattaatgta agttagcgcg aattgatctg gtttgacagc ttatcatcga ctgcacggtg
     4081 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
     4141 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     4201 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatccg
     4261 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagcgccgct
     4321 gagaaaaagc gaagcggcac tgctctttaa caatttatca gacaatctgt gtgggcactc
     4381 gaccggaatt atcgattaac tttattatta aaaattaaag aggtatatat taatgtatcg
     4441 attaaataag gaggaataaa ccatggcagc cgggaggtgc atagtgagaa catacgagaa
     4501 cgcgcaggag ctcaaaaagg agatcggcgc ggcgtttcga aaatacattg cggagttcga
     4561 cgacattccg gaagccctga aagacaagcg catcgacgag gtcgagagga ccccggcgga
     4621 aaacctcgcc tatcaggttg gttggaccac cctgctgctg caatgggagg atcgcgagcg
     4681 gaagggcctt ccggtgcgaa cgccgtcgga cgagttcaag tggaaccagc tcggaaagct
     4741 gtaccagtgg ttcaccgaca cctacgccca cctttcgctg cgggagctga agggcat
//
