LOCUS       sequence-318836-        2151 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 2151)
  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 2151)
  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..2151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(199..233)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(310..327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(481..1069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(561..580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1243..2034)
                     /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 actcgagtag ggataacagg gtaatagatc taagcttctg caggtcgact ctagagaatt
       61 caccttactc gagtttggat ccgagttttt gttcgggccc aagcttcaaa aaaaataatg
      121 cgaacagcgc acggcgtatc tacaagagta gaaattcgtc acactacgtc tgaacgtcga
      181 tctacaagag tagaaattac tagtattata cctaggactg agctagctgt caaggatcca
      241 gcatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
      301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      901 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
      961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1141 catgagatta tcaaaaagga tcttcaccta gatcctttac gcgttcgcgg aacttcggaa
     1201 taggaactta tgagctcagc caatcgactg gcgagcggca tcttatttgc cgactacctt
     1261 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
     1321 gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
     1381 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     1441 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     1501 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     1561 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     1621 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
     1681 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     1741 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     1801 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     1861 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc
     1921 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac
     1981 gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc tcatgacatt
     2041 gcactccacc gctgatgaca tcagtcgatc atagcacgat caacggcact gttgcaaata
     2101 gtcggtggtg ataaacttat catccccttt tgcttatgga gctgcacatg a
//
