LOCUS       sequence-318838-        3332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPR-Cas12a genome editing in E. coli. Contains a 
            constituitively expressed CRISPR array targeting the ahpC gene in E.
            coli and donor DNA to insert an rfp translational fusion in the ahpC
            gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3332)
  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 3332)
  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-ahpC-rfp
FEATURES             Location/Qualifiers
     source          1..3332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             143..934
                     /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"
     rep_origin      1108..1696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1597..1616
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1850..1867
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1944..1978
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     terminator      2085..2156
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2172..2199
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2418..3095
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 attaccctgt tatccctact cgagttcatg tgcagctcca taagcaaaag gggatgataa
       61 gtttatcacc accgactatt tgcaacagtg ccgttgatcg tgctatgatc gactgatgtc
      121 atcagcggtg gagtgcaatg tcatgaggga agcggtgatc gccgaagtat cgactcaact
      181 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
      241 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
      301 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
      361 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
      421 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
      481 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
      541 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
      601 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
      661 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
      721 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
      781 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
      841 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
      901 gaaaggcgag atcaccaagg tagtcggcaa ataagatgcc gctcgccagt cgattggctg
      961 agctcataag ttcctattcc gaagttccgc gaacgcgtaa aggatctagg tgaagatcct
     1021 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1081 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1141 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1201 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     1261 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1321 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1381 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1441 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1501 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1561 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1621 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1681 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1741 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1801 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1861 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1921 ttcacaccgc atatgctgga tccttgacag ctagctcagt cctaggtata atactagtaa
     1981 tttctactct tgtagatacc aaccaggtcc agagacggag tttttttgaa gcttgggccc
     2041 gaacaaaaac tcggatccaa actcgagtaa ggatctccag gcatcaaata aaacgaaagg
     2101 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     2161 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatacctag ggtacgatgt
     2221 ccttgattaa caccaaaatt aaacctttta aaaaccaggc attcaaaaac ggcgaattca
     2281 tcgaaatcac cgaaaaagat accgaaggcc gctggagcgt cttcttcttc tacccggctg
     2341 actttacttt cgtatgcccg accgaactgg gtgacgttgc tgaccactac gaagaactgc
     2401 agaaactggg cgtagacatg gcgagtagcg aagacgttat caaagagttc atgcgtttca
     2461 aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag
     2521 gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc
     2581 cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac
     2641 acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac
     2701 gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag
     2761 acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg
     2821 ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg
     2881 gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg
     2941 ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa
     3001 ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg
     3061 aacgtgctga aggtcgtcac tccaccggtg cttaaatttc cttcgtcttt cacgccatag
     3121 cggcgttggc gtcgcccgct caccccggtc acttacttgt gtaagctccc ggggattcac
     3181 aggctagccg ccttgctctg acgcgaaata cttcggaaat tcacctaatt cttcgggtgc
     3241 tgcggcaccc gattttcttc cccgcaccat gatgcaagct gcatccaggt agccgtgaat
     3301 tctctagagt cgacctgcag aagcttagat ct
//
