LOCUS       sequence-336412-        2243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of sgRNA without donor DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2243)
  AUTHORS   Okano K, Sato Y, Hizume T, Honda K
  TITLE     Genome editing by miniature CRISPR/Cas12f1 enzyme in Escherichia 
            coli.
  JOURNAL   J Biosci Bioeng. 2021 Aug;132(2):120-124. doi: 
            10.1016/j.jbiosc.2021.04.009. Epub 2021 May 19.
  PUBMED    34023220
REFERENCE   2  (bases 1 to 2243)
  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"; doi: 
            "10.1016/j.jbiosc.2021.04.009"; journalName: "J Biosci Bioeng"; 
            date: "2021-08"; volume: "132"; issue: "2"; pages: "120-124"
COMMENT     Sequence Label: pTarget12f1
FEATURES             Location/Qualifiers
     source          1..2243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        192..226
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             634..1425
                     /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      1599..2187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2088..2107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      181 atgctggatc cttgacagct agctcagtcc taggtataat actagtcttc actgataaag
      241 tggagaaccg cttcaccaaa agctgtccct taggggatta gaacttgagt gaaggtgggc
      301 tgcttgcatc agcctaatgt cgagaagtgc tttcttcgga aagtaaccct cgaaacaaat
      361 tcatttttcc tctccaattc tgcacaagaa agttgcagaa cccgaataga cgaatgaagg
      421 aatgcaactg caccgtgtct agctagcttt tttttgaatt ctctagagtc gacctgcaga
      481 agcttagatc tattaccctg ttatccctac tcgagttcat gtgcagctcc ataagcaaaa
      541 ggggatgata agtttatcac caccgactat ttgcaacagt gccgttgatc gtgctatgat
      601 cgactgatgt catcagcggt ggagtgcaat gtcatgaggg aagcggtgat cgccgaagta
      661 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
      721 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
      781 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
      841 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
      901 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
      961 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     1021 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     1081 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     1141 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     1201 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     1261 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     1321 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     1381 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataagatgc cgctcgccag
     1441 tcgattggct gagctcataa gttcctattc cgaagttccg cgaacgcgta aaggatctag
     1501 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1561 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1621 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1681 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1741 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1801 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1861 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1921 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1981 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2041 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2101 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2161 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2221 gccttttgct ggccttttgc tca
//
