LOCUS       sequence-124217-        2584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a toehold mutant of the circRAJ31 riboregulator encased in
            a permuted intron exon ribozyme, which circularises the 
            riboregulator. This mutant does not activate cisRAJ31..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2584)
  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 2584)
  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: circRAJ31-Toehold_mutant
FEATURES             Location/Qualifiers
     source          1..2584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6..22
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    222..240
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(792..808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(792..808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      856..887
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        888..990
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(991..1059,1060..1851)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGTRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1209..1228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      1923..1950
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1962..2549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2042..2061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgtgtaaa acgacggcca gtttatctag tcagcttgat tctagctgat cgtggaccgg
       61 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcgtcctgaa
      121 tgatatgcga ccgccggagg gttgcgtttg agacgggcga cagatcgaca ctgctcgatc
      181 cgctcgcacc gaattctccc tatcagtgat agagattgac atccctatca gtgatagaga
      241 tactgagcac tctagaggtt ctacataaat gcctaacgac tatccctttg gggagtaggg
      301 tcaagtgact cgaaacgata gacaacttgc tttaacaagt tggagatata gtctgctctg
      361 catggtgaca tgcagctgga tataattccg gggtaagatt aacgacctta tctgaacata
      421 atgctaccgt ttaatattcc taggggatca atattaagct ttaccgacac cccatccgca
      481 attaagtccc tatccgctgt cactgtattg gtctgctagc atgttttctt gggttaattg
      541 aggcctgagt ataaggtgac ttatacttgt aatctatcta aacggggaac ctctctagta
      601 gacaatcccg tgctaaattg taggactcaa ttgatcctta gcgaaagcta aggatttttt
      661 ttctgcaggg atcgacgaga gcagcgcgac tggatcagtt ctggacgagc gagctgtcgt
      721 ccgacccgtg atcttacggc attatacgta tgatcggtcc acgatcagct agattatcta
      781 gtcagcttga tgtcatagct gtttcctgag gctcaatact gaccatttaa atcatacctg
      841 acctccatag cagaaagtca aaagcctccg accggaggct tttgacttga tcggcacgta
      901 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
      961 tcgagatttt caggagctaa ggaagctaaa atgagtattc aacatttccg tgtcgccctt
     1021 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1081 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1141 agcggtaaga tccttgagag tttacgcccc gaagaacgtt ttccaatgat gagcactttt
     1201 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1261 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1321 ctcacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1381 actgcggcca acttacttct ggcaacgatc ggaggaccga aggagctaac cgcttttttg
     1441 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1501 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1561 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1621 gcggataaag ttgcaggatc acttctgcgc tcggccctcc cggctggctg gtttattgct
     1681 gataaatctg gagccggtga gcgtgggact cgcggtatca ttgcagcact ggggccagat
     1741 ggtaagccct cccgcatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1801 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta atgagggccc
     1861 aaatgtaatc acctggctca ccttcgggtg ggcctttctt gaggacctaa atgtaatcac
     1921 ctggctcacc ttcgggtggg cctttctgcg ttgctggcgt ttttccatag gctccgcccc
     1981 cctgacgagc atcacaaaaa tcgatgctca agtcagaggt ggcgaaaccc gacaggacta
     2041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2401 agaacagtat ttggtatctg cgctctgctg aagccagtta cctcggaaaa agagttggta
     2461 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2521 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gattttctac cgaagaaagg
     2581 ccca
//
