LOCUS       sequence-441239-        3111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pORI28 derivative with lacZ and upp gene inserted into the backbone;
            for generating gene-specific knockouts..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3111)
  AUTHORS   Zhu M, Frank MW, Radka CD, Jeanfavre S, Tse MW, Pacheco JA, Pierce 
            K, Deik A, Xu J, Hussain S, Hussain FA, Xulu N, Khan N, Pillay V, 
            Dong KL, Ndung'u T, Clish CB, Rock CO, Blainey PC, Bloom SM, Kwon DS
  TITLE     Vaginal Lactobacillus fatty acid response mechanisms reveal a novel 
            strategy for bacterial vaginosis treatment.
  JOURNAL   bioRxiv [Preprint]. 2023 Dec 30:2023.12.30.573720. doi: 
            10.1101/2023.12.30.573720.
  PUBMED    38234804
REFERENCE   2  (bases 1 to 3111)
  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: "bioRxiv 
            [Preprint]. 2023 Dec 30:2023.12.30.573720. doi: 
            10.1101/2023.12.30.573720."
COMMENT     Sequence Label: pMZ7
FEATURES             Location/Qualifiers
     source          1..3111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(175..197)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(292..615)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     297..316
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     510..532
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     524..541
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     525..541
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    542..598
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(611..627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(611..627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(624..646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    635..651
                     /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(659..665,666..683,684..689))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    704..725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(842..859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             1357..1368
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ataattgtta atttgtgctg aatcaaaggg tgatccaaaa ctacaaactt tcccatatat
       61 gcctcctatt aacaaataac ttaattcatt aaatatttta atcaaaaacg caaagggttt
      121 aaagcccttt gcgttttatt ttttatttat tcaaatgatg gccgccggtg aaaacctctg
      181 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
      241 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
      301 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
      361 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
      421 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
      481 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      541 tgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg
      601 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      661 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      721 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      781 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      841 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      901 caaaggcggt aatacggtta tccacagaat caggggataa cgcagctaga actagcgatt
      961 ctgaaatcac catttaaaaa actccaatca aataatttta taaagttagt gtatcacttt
     1021 gtaatcataa aaacaacaat aaagctactt aaatatagat ttataaaaaa cgttggcgaa
     1081 aacgttggcg attcgttggc gattgaaaaa ccccttaaac ccttgagcca gttgggatag
     1141 agcgtttttg gcacaaaaat tggcactcgg cacttaatgg ggggtcgtag tacggaagca
     1201 aaattcgctt cctttccccc catttttttc caaattccaa atttttttca aaaattttcc
     1261 agcgctaccg ctcggcaaaa ttgcaagcaa tttttaaaat caaacccatg agggaatttc
     1321 attccctcat actcccttga gcctcctcca accgaaatag aagggcgctg cgcttattat
     1381 ttcattcagt catcggcttt cataatctaa cagacaacat cttcgctgca aagccacgct
     1441 acgctcaagg gcttttacgc tacgataacg cctgttttaa cgattatgcc gataactaaa
     1501 cgaaataaac gctaaaacgt ctcagaaacg attttgagac gttttaataa aaaatcgcta
     1561 gtccgaggcc tcgacccgat tcacaaaaaa taggcacacg aaaaacaagt taagggatgc
     1621 agtttatgca tcccttaact tacttattaa ataatttata gctattgaaa agagataaga
     1681 attgttcaaa gctaatattg tttaaatcgt caattcctgc atgttttaag gaattgttaa
     1741 attgattttt tgtaaatatt ttcttgtatt ctttgttaac ccatttcata acgaaataat
     1801 tatacttttg tttatctttg tgtgatattc ttgatttttt tctacttaat ctgataagtg
     1861 agctattcac tttaggttta ggatgaaaat attctcttgg aaccatactt aatatagaaa
     1921 tatcaacttc tgccattaaa agtaatgcca atgagcgttt tgtatttaat aatcttttag
     1981 caaacccgta ttccacgatt aaataaatct cattagctat actatcaaaa acaattttgc
     2041 gtattatatc cgtacttatg ttataaggta tattaccata tattttatag gattggtttt
     2101 taggaaattt aaactgcaat atatccttgt ttaaaacttg gaaattatcg tgatcaacaa
     2161 gtttattttc tgtagttttg cataatttat ggtctatttc aatggcagtt acgaaattac
     2221 acctctttac taattcaagg gtaaaatggc cttttcctga gccgatttca aagatattat
     2281 catgttcatt taatcttata tttgtcatta ttttatctat attatgtttt gaagtaataa
     2341 agttttgact gtgttttata tttttctcgt tcattataac cctctttaat ttggttatat
     2401 gaattttgct tattaacgat tcattataac cacttatttt ttgtttggtt gataatgaac
     2461 tgtgctgatt acaaaaatac taaaaatgcc catatttttt cctccttata aaattagtat
     2521 aattatagca cggtcgatta cttagtaccg aagagtctgt caccggcatc tcccaaacca
     2581 gggaaaatgt aaccattgtc cagcaattta tcatcttctg atgcagcata aatatcaaca
     2641 tcaggatttt cttcttggac tgccttaata ccttctggag ctgctactaa aactgccaat
     2701 ttaatatcct taacaccacg cttctttaat gcagcgatag cgtcattagc agaaccacca
     2761 gtagcaagca ttggatcaac aactaaacat tcacgttcag caatatcctt tggagcttta
     2821 aagaaatatt catgcggctt caaagtttct tcatcacgat acataccaat aacagcaatc
     2881 ttagctgatg gtaccatctc aagaacaccg ttaagcattc ccatacctgc acgcaaaatc
     2941 ggcacaattg tcaatttctt gccggcgatt tctttttgga ctgtcttacc cattggtgtt
     3001 tcaatttcaa catcttcaag tggtaaatct ctagtaattt cataggtcat taatccaccg
     3061 atttcaccaa caattctacg aaattcgttt gaacctgtgt ccttgcgacg a
//
