LOCUS       sequence-437240-        2584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible CRISPRi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2584)
  AUTHORS   Mayers JR, Varon J, Zhou RR, Daniel-Ivad M, Beaulieu C, Bholse A, 
            Glasser NR, Lichtenauer FM, Ng J, Vera MP, Huttenhower C, Perrella 
            MA, Clish CB, Zhao SD, Baron RM, Balskus EP
  TITLE     A metabolomics pipeline highlights microbial metabolism in 
            bloodstream infections.
  JOURNAL   Cell. 2024 Jun 11:S0092-8674(24)00579-8. doi: 
            10.1016/j.cell.2024.05.035.
  PUBMED    38885650
REFERENCE   2  (bases 1 to 2584)
  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"; journalName: "Cell. 2024 
            Jun 11:S0092-8674(24)00579-8. doi: 10.1016/j.cell.2024.05.035."
COMMENT     Sequence Label: pgRNA-bacteria_speG_M15
FEATURES             Location/Qualifiers
     source          1..2584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..39
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        60..135
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             156..185
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             201..218
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(274..291)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      444..490
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(543..560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(714..1302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(794..813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1473..2264,2265..2333))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2096..2115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2334..2438)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2506..2524
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2528..2571)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 atctttgaca gctagctcag tcctaggtat aatactagtt taaaggtgat tctacaccag
       61 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
      121 gcaccgagtc ggtgcttttt ttgaagcttg ggcccgaaca aaaactcatc tcagaagagg
      181 atctgaatag cgccgtcgac catcatcatc atcatcattg agtttaaacg gtctccagct
      241 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
      301 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
      361 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
      421 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      481 ttcgttttat ctgttgtttg tcggtgaact ggatccttac tcgagtctag actgcaggct
      541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccac
      721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2461 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2521 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattct
     2581 aaag
//
