LOCUS       sequence-461663-        2356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ISFba1 guide RNA targeting maeB site controled by 
            PJ23119.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2356)
  AUTHORS   Zhou X, Yang S, Sun B, Dong F, Yin M, Jiang Y, Huang Z, Yang S
  TITLE     Implementation of RAGATH RNA-associated DNA Endonucleases as Genome 
            Editing Tool in Escherichia coli.
  JOURNAL   Biotechnol J. 2025 Mar;20(3):e70005. doi: 10.1002/biot.70005.
  PUBMED    40111000
REFERENCE   2  (bases 1 to 2356)
  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: "Biotechnol 
            J."; date: "2025-03"; volume: "20(3):e70005. doi"; pages: " 
            10.1002/biot.70005"
COMMENT     Sequence Label: pISFba1reRNA
FEATURES             Location/Qualifiers
     source          1..2356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..39
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     ncRNA           247..314
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     primer_bind     complement(353..371)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      367..414
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             560..1351
                     /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      1525..2113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2014..2033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2267..2284
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atccttgaca gctagctcag tcctaggtat aatactagtc taaaacgcaa acgtaagtat
       61 gtaccgaagg ctattttcgg gaatttacga ctatggagtg tacaagaact tgtgagtaga
      121 tatgatttcg gtcaatccaa agcatacacg atgaaatagt aagtaatgtt cgtgagaact
      181 tacattatct cgatgtgagc atatttaatc acattttgag tggcagctga tggtccatgt
      241 ctgttaggcc ggcatggtcc cagcctcctc gctggcgccg gctgggcaac atgcttcggc
      301 atggcgaatg ggactaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
      361 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
      421 ggaggaacta tatccgtcga gttcatgtgc agctccataa gcaaaagggg atgataagtt
      481 tatcaccacc gactatttgc aacagtgccg ttgatcgtgc tatgatcgac tgatgtcatc
      541 agcggtggag tgcaatgtca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
      601 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
      661 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
      721 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
      781 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
      841 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
      901 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
      961 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     1021 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     1081 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     1141 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     1201 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     1261 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     1321 aggcgagatc accaaggtag tcggcaaata agatgccgct cgccagtcga ttggctgagc
     1381 tcataagttc ctattccgaa gttccgcgaa cgcgtaaagg atctaggtga agatcctttt
     1441 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1501 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1561 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1621 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1681 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1741 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1801 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1861 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1921 agaaagcgcc acgcttcccg aagggaggaa ggcggacagg tatccggtaa gcggcagggt
     1981 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2041 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2101 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2161 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2221 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2281 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2341 acaccgcata tgctgg
//
