LOCUS       sequence-296088-        2152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPR-Cas12a genome editing in E. coli for the insertion of 
            large DNA fragments at the lacZ locus. Contains a constitutively 
            expressed CRISPR array targeting the lacZ gene . No cargo DNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2152)
  AUTHORS   Jervis AJ, Hanko EKR, Dunstan MS, Robinson CJ, Takano E, Scrutton NS
  TITLE     A plasmid toolset for CRISPR-mediated genome editing and CRISPRi 
            gene regulation in Escherichia coli.
  JOURNAL   Microb Biotechnol. 2021 Mar 12. doi: 10.1111/1751-7915.13780.
  PUBMED    33710766
REFERENCE   2  (bases 1 to 2152)
  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: "Microb 
            Biotechnol. 2021 Mar 12. doi: 10.1111/1751-7915.13780."
COMMENT     Sequence Label: pTF-lacZ
FEATURES             Location/Qualifiers
     source          1..2152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(193..227)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(304..321)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(475..1063)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(555..574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1238..2029)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gtagggataa cagggtaata gatctaagct tctgcaggtc gactctagag aattcacctt
       61 actcgagttt ggatccgagt ttttgttcgg gcccaagctt caaaaaaaat aatgcgaaca
      121 gcgcacggcg tatctacaag agtagaaatt cgtcacacta cgtctgaacg tcgatctaca
      181 agagtagaaa ttactagtat tatacctagg actgagctag ctgtcaagga tccagcatat
      241 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
      301 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
      361 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
      421 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      481 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      541 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      601 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      661 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      721 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      781 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      841 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      901 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      961 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1021 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1081 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1141 attatcaaaa aggatcttca cctagatcct ttacgcgttc gcggaacttc ggaataggaa
     1201 cttcatgagc tcagccaatc gactggcgag cggcatctta tttgccgact accttggtga
     1261 tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat
     1321 cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg
     1381 gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg
     1441 cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg
     1501 gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa
     1561 ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt
     1621 ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa
     1681 tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa
     1741 tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag
     1801 gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc
     1861 ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatccactg
     1921 cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg atgacgccaa
     1981 ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcatg acattgcact
     2041 ccaccgctga tgacatcagt cgatcatagc acgatcaacg gcactgttgc aaatagtcgg
     2101 tggtgataaa cttatcatcc ccttttgctg atggagctgc acatgaactc ga
//
