LOCUS       sequence-244554-        3894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes an RNA nanostructure with MG, PP7, TAR and S1m RNA aptamers.
            Transcription via T7 RNAP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3894)
  AUTHORS   Schwarz-Schilling M, Dupin A, Chizzolini F, Krishnan S, Mansy SS, 
            Simmel FC
  TITLE     Optimized Assembly of a Multifunctional RNA-Protein Nanostructure in
            a Cell-Free Gene Expression System.
  JOURNAL   Nano Lett. 2018 Apr 11;18(4):2650-2657. doi: 
            10.1021/acs.nanolett.8b00526. Epub 2018 Mar 27.
  PUBMED    29564885
REFERENCE   2  (bases 1 to 3894)
  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"; doi: 
            "10.1021/acs.nanolett.8b00526"; journalName: "Nano Lett"; date: 
            "2018-04-11- 11"; volume: "18"; issue: "4"; pages: "2650-2657"
COMMENT     Sequence Label: pSB4k5_RNAnano (r-plasmid)
FEATURES             Location/Qualifiers
     source          1..3894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      589..811
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(1303..1397)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(1553..1582)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    1629..1650
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     1663..1682
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1663..1681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2057..2075)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2071..2118
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2182..2213
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2312..3127)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3035..3054
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3119..3138
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(join(3485..3894,1..541))
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg
       61 ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt
      121 atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga
      181 aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg
      241 catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag
      301 ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga
      361 tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag
      421 ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct
      481 ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca
      541 tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat
      601 gataattaca tgtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg
      661 ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt
      721 tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa
      781 aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca
      841 aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct
      901 taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc
      961 aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca
     1021 gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc
     1081 ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt
     1141 ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca
     1201 cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt
     1261 atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa
     1321 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     1381 atctatcaac aggagtccaa gcgagctcgt aaacttggtc tgacagctct agctccggca
     1441 aaaaaacggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc
     1501 gcgtttgcca cctgacgtct aagaaaagga atattcagca atttgcccgt gccgaagaaa
     1561 ggcccacccg tgaaggtgag ccagtgagtt gattgctacg taattagtta gttagccctt
     1621 agtgactcga attcgcggcc gcttctagag cacaggccac agtaatacga ctcactatag
     1681 ggttgtcatg tgtatgttgg gggatcccga ctggcgagag ccaggtaacg aatggatccc
     1741 ccacatactt tgttgatccc gcaggattcg gctcgtgtag ctcattagct ccgagccgag
     1801 tcctcgaata cgagctgggc acagaagata tggcttcgtg cccaggaggt gttcgcactt
     1861 ctctcgtgtt cgattgtggg atcaatcatg gcaaatgcgg ccgccgacca gaatcatgca
     1921 agtgcgtaag atagtcgcgg gtcggtggtc gcattactta gcataacccc gcggggcctc
     1981 ttcgggggtc tcgcggggtt ttttgctgaa acggctgcta acaaagcccg aaaggaagct
     2041 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     2101 gtcttgaggg gttttttgct gaaaggagga actacactgt ctgcaggagt cactaagggt
     2161 tagttagtta gattagcaga aagtcaaaag cctccgaccg gaggcttttg actaaaactt
     2221 cccttggggt tatcattggg gctcactcaa aggcggtaat cagataaaaa aaatccttag
     2281 ctttcgctaa ggatgatttc tgctagtatt attagaaaaa ctcatcgagc atcaaatgaa
     2341 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     2401 atgaaggaga aaactcaccg aggcagttcc aaagaatggc aaggtcctgg taacggtctg
     2461 cgattccgac ccgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     2521 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aagagcttgt
     2581 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     2641 catcaaccaa accgttattc atgcgtgatt gcgcctgagc aagacgaaat acacgatcgc
     2701 tgttaaaagg acaattacaa acaggaatcg aatgtaaccg gcgcaggaac acggccagcg
     2761 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaag gctgttttcc
     2821 caggaatcgc ggtggtgagt aaccacgcat catcaggagt acggataaaa tgcttgatgg
     2881 tcgggagagg cataaactcc gtcagccagt tgagacggac catctcatct gtaacatcat
     2941 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     3001 agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     3061 aatcagcgtc catgttggag tttaagcgcg gacgggagca agacgtttcc cgttgaatat
     3121 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     3181 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt
     3241 gaataaatcg aacttttgct gagttgaagg atcagctcta gtagttacat tgtcgatctg
     3301 ttcatggtga acagctttga atgcaccaaa aactcgtaaa agctctgatg tatctatctt
     3361 ttttacaccg ttttcatctg tgcatatgga cagttttccc tttgatatgt aacggtgaac
     3421 agttgttcta cttttgtttg ttagtcttga tgcttcactg atagatacaa gagccataag
     3481 aacctcagat ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg
     3541 tgagccatga gaacgaacca ttgagatcat acttactttg catgtcactc aaaaattttg
     3601 cctcaaaact ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc
     3661 cgttatgtag gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta
     3721 tctggttgtt ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa
     3781 tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt
     3841 ttaaatcttt acttattggt ttcaaaaccc attggttaag ccttttaaac tcat
//
