LOCUS       sequence-458204-        2370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ISEre1 guide RNA targeting maeB site controled by 
            PJ23119.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2370)
  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 2370)
  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: pISEre1reRNA
FEATURES             Location/Qualifiers
     source          1..2370
                     /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           244..311
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     primer_bind     complement(367..385)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      381..428
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             574..1365
                     /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      1539..2127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2028..2047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2281..2298
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atccttgaca gctagctcag tcctaggtat aatactagta cgcaaacgta agtatgtact
       61 gcgggctatc gcaggaattt acgactgtgg agtgtacacg aacttgtgag tagcgtattg
      121 tttacaatcg ccaaagcata cacatcgaag cagtaagaag tatccgcaag gactttaatt
      181 tctcgatgtg tttgagtata tttgaacaca ttttgagtgg cagctgatgg tccatgtctg
      241 ttaggccggc atggtcccag cctcctcgct ggcgccggct gggcaacatg cttcggcatg
      301 gcgaatggga cctcgaggat ccggctgcta acaaagcccg aaaggaagct gagttggctg
      361 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
      421 gttttttgct gaaaggagga actatatccg tcgagttcat gtgcagctcc ataagcaaaa
      481 ggggatgata agtttatcac caccgactat ttgcaacagt gccgttgatc gtgctatgat
      541 cgactgatgt catcagcggt ggagtgcaat gtcatgaggg aagcggtgat cgccgaagta
      601 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
      661 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
      721 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
      781 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
      841 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
      901 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
      961 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     1021 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     1081 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     1141 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     1201 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     1261 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     1321 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataagatgc cgctcgccag
     1381 tcgattggct gagctcataa gttcctattc cgaagttccg cgaacgcgta aaggatctag
     1441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1681 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2221 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2281 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     2341 ctgtgcggta tttcacaccg catatgctgg
//
