LOCUS       sequence-467174-        4229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modular RENDR, GFP output, targets synthetic target.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4229)
  AUTHORS   Gambill L, Staubus A, Mo KW, Ameruoso A, Chappell J
  TITLE     A split ribozyme that links detection of a native RNA to orthogonal 
            protein outputs.
  JOURNAL   Nat Commun. 2023 Feb 1;14(1):543. doi: 10.1038/s41467-023-36073-3.
  PUBMED    36725852
REFERENCE   2  (bases 1 to 4229)
  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: "Nat 
            Commun."; date: "2023-02-1"; volume: "14(1):543. doi"; pages: " 
            10.1038/s41467-023-36073-3"
COMMENT     Sequence Label: pJEC779
FEATURES             Location/Qualifiers
     source          1..4229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(566..657)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(705..1443)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     promoter        1702..1736
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     promoter        complement(1925..2029)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2097..2115
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2119..2162)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        2180..2214
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(2711..2713,2714..3427)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2711..3424
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2741..2769)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3364..3384
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3496..3513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3569..3586)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3739..3785
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(4003..4229,1..565))
                     /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"
ORIGIN
        1 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
       61 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
      121 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
      181 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa
      241 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
      301 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt
      361 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
      421 caatatcact gtgtggctta aggccgccat ccactgcgga gccgtacaaa tgtacggcca
      481 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
      541 cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga agcatttatc
      601 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
      661 ctagctcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata
      721 caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag
      781 ggccgcgccg gtggcgtttt tccataggct ccgccatcct gccagagttc acataaacag
      841 acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg
      901 cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct
      961 ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag
     1021 tgtggcgctt tctcatagct aacacactgg tatctcggct cggtgtaggt cgttcgctcc
     1081 aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac
     1141 tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt
     1201 aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc
     1261 caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac
     1321 cacggttaag cagttcacca actgacttaa ccttcgatca aaccacctcc caaggtggtt
     1381 ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1441 atcttttcta ctgaaccgct ctagatttca gtgcaattta tctcttcaaa tgtagcacct
     1501 gaagtcagcc caatgcgata taagttgtaa ttctcatgtt agtcatgcac cgcgcccacc
     1561 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     1621 gagtgagcta acttacaccc gcagttccag tacggttcca ttaattgcgt tgcgctgctt
     1681 ccggctgaat tctaaagatc tttgacagct agctcagtcc taggtataat actagtacgt
     1741 cgactctcga gtgagattgt tgacggtacc gtattttgga tctaggagga atgacctaaa
     1801 tagcaaggat cattcattct gcccacaatg acgaatctag gtcggtcaca ttaggtcctc
     1861 gaagttgcaa cggcgtcccc acatcgttga gtggcaaagc ccgccgaaag gcgggctttt
     1921 ttttactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1981 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     2041 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     2101 ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt
     2161 tctggaattc taaagatctt tgacagctag ctcagtccta ggtataatac tagtggatat
     2221 ataacctgcg caagaggccg gacgtattat gagactctct taccgttagg tcaggtcttg
     2281 ttcggcactc aatgtctgcc tgatcctagt tacctttggg tcaaaaagtt atcaggcatg
     2341 cacctggtag ctagtcttta aaccaataga ttgcatcggt ttaaaaggca agaccgtcaa
     2401 attgcgggaa aggggtcaac agccgttcag taccaagtct caggggaaac tttgagatgg
     2461 ccttgcaaag ggtatggtaa taagctgacg gacatggtcc taaccacgca gccaagtcct
     2521 aagtcaacag atcttctgtt gatatggatg cagttcacag actaaatgtc ggtcggggaa
     2581 gatgtattct tctcataaga tatagtcgga cctctcctta atgggagcta gcggatgaag
     2641 tgatgcaaca ctggagccgc tgggaactaa tttgtatgcg aaagtatatt gattagtttt
     2701 ggagtactcg atgagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     2761 attagatggt gatgttaatg ggcacaaatt ttctgtccgt ggagagggtg aaggtgatgc
     2821 tacaaacgga aaactcaccc ttaaatttat ttgcactact ggaaaactac ctgttccgtg
     2881 gccaacactt gtcactactc tgacctatgg tgttcaatgc ttttcccgtt atccggatca
     2941 catgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac
     3001 tatatctttc aaagatgacg ggacctacaa gacgcgtgct gaagtcaagt ttgaaggtga
     3061 tacccttgtt aatcgtatcg agttaaaggg tattgatttt aaagaagatg gaaacattct
     3121 tggacacaaa ctcgagtaca actttaactc acacaatgta tacatcacgg cagacaaaca
     3181 aaagaatgga atcaaagcta acttcaaaat tcgccacaac gttgaagatg gttccgttca
     3241 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     3301 caaccattac ctgtcgacac aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca
     3361 catggtcctt cttgagtttg taactgctgc tgggattaca catggcatgg atgagctcta
     3421 caaataatat gggatctgaa gcttgggccc taacaaaaac tcatctcaga agaggatctg
     3481 aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacggtctc cagcttggct
     3541 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     3601 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     3661 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     3721 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     3781 tttatctgtt gtttgtcggt gaactggatc cttactcgag tctagactgc agctgaaacc
     3841 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     3901 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcatcgtg
     3961 gccggatctt gcggccactc ggcttgaacg aattgttaga cattatttgc cgactacctt
     4021 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
     4081 gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
     4141 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     4201 tactgcgctg taccaaatgc gggacaacg
//
