LOCUS       sequence-124224-        2491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RAJ31 riboregulator encased in a permuted intron exon 
            ribozyme, which circularises the riboregulator. This mutant is 
            deficient in circularisation. Under control of a T7 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2491)
  AUTHORS   Rostain W, Shen S, Cordero T, Rodrigo G, Jaramillo A
  TITLE     Engineering a Circular Riboregulator in Escherichia coli.
  JOURNAL   Biodes Res. 2020 Sep 12;2020:1916789. doi: 10.34133/2020/1916789. 
            eCollection 2020.
  PUBMED    37849901
REFERENCE   2  (bases 1 to 2491)
  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: "Biodes 
            Res."; date: "2020-09-12"; volume: "2020:1916789. doi"; pages: " 
            10.34133/2020/1916789. eCollection 2020"
COMMENT     Sequence Label: pT7_circRAJ31_C873U
FEATURES             Location/Qualifiers
     source          1..2491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      7..65
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(89..106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(260..848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(340..359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1031..1125)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1146..1805)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1720..1739
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1967..1985
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1989..2032)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     2041..2060
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2041..2059
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2444..2491
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ctgcagtccg gcaaaaaaac ggcaaggtgt caccaccctg ccctttttct ttaaaaccga
       61 aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
      121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      241 aggccgcgtt gctggcgttt ttccacaggc tccgcccccc tgacgagcat cacaaaaatc
      301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      661 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct
     1021 cgaggcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     1081 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata
     1141 tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     1201 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     1261 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     1321 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     1381 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     1441 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     1501 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     1561 agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga
     1621 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     1681 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     1741 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     1801 tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     1861 gatcttattt cattatggtg aaagttggaa cctcttacgt gcccgatcaa ctcgagtgcc
     1921 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     1981 gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc
     2041 taatacgact cactataggg tctagaggtt ctacataaat gcctaacgat tatccctttg
     2101 gggagtaggg tcaagtgact cgaaacgata gacaacttgc tttaacaagt tggagatata
     2161 gtctgctctg catggtgaca tgcagctgga tataattccg gggtaagatt aacgacctta
     2221 tctgaacata atgctaccgt ttaatattcc taggggatca atattaagct ttaccgtcgt
     2281 ctcatccgca attaagtccc tatccgctgt cactgtattg gtctgctagc atgttttctt
     2341 gggttaattg aggcctgagt ataaggtgac ttatacttgt aatctatcta aacggggaac
     2401 ctctctagta gacaatcccg tgctaaattg taggactcaa ttgctagcat aaccccttgg
     2461 ggcctctaaa cgggtcttga ggggtttttt g
//
