LOCUS       sequence-249161-        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"
     terminator      complement(1..30)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    77..98
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     111..130
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        111..129
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(505..523)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      519..566
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      630..661
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(760..1575)
                     /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     1483..1502
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1567..1586
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1933..2883)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFGIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2931..3153
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3645..3739)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 cgaagaaagg cccacccgtg aaggtgagcc agtgagttga ttgctacgta attagttagt
       61 tagcccttag tgactcgaat tcgcggccgc ttctagagca caggccacag taatacgact
      121 cactataggg ttgtcatgtg tatgttgggg gatcccgact ggcgagagcc aggtaacgaa
      181 tggatccccc acatactttg ttgatcccgc aggattcggc tcgtgtagct cattagctcc
      241 gagccgagtc ctcgaatacg agctgggcac agaagatatg gcttcgtgcc caggaggtgt
      301 tcgcacttct ctcgtgttcg attgtgggat caatcatggc aaatgcggcc gccgaccaga
      361 atcatgcaag tgcgtaagat agtcgcgggt cggtggtcgc attacttagc ataaccccgc
      421 ggggcctctt cgggggtctc gcggggtttt ttgctgaaac ggctgctaac aaagcccgaa
      481 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
      541 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tacactgtct gcaggagtca
      601 ctaagggtta gttagttaga ttagcagaaa gtcaaaagcc tccgaccgga ggcttttgac
      661 taaaacttcc cttggggtta tcattggggc tcactcaaag gcggtaatca gataaaaaaa
      721 atccttagct ttcgctaagg atgatttctg ctagtattat tagaaaaact catcgagcat
      781 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
      841 tttctgtaat gaaggagaaa actcaccgag gcagttccaa agaatggcaa ggtcctggta
      901 acggtctgcg attccgaccc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
      961 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     1021 gagcttgtgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     1081 atcactcgca tcaaccaaac cgttattcat gcgtgattgc gcctgagcaa gacgaaatac
     1141 acgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgtaaccggc gcaggaacac
     1201 ggccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaaggc
     1261 tgttttccca ggaatcgcgg tggtgagtaa ccacgcatca tcaggagtac ggataaaatg
     1321 cttgatggtc gggagaggca taaactccgt cagccagttg agacggacca tctcatctgt
     1381 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     1441 cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     1501 cccatataaa tcagcgtcca tgttggagtt taagcgcgga cgggagcaag acgtttcccg
     1561 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     1621 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     1681 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagctctagt agttacattg
     1741 tcgatctgtt catggtgaac agctttgaat gcaccaaaaa ctcgtaaaag ctctgatgta
     1801 tctatctttt ttacaccgtt ttcatctgtg catatggaca gttttccctt tgatatgtaa
     1861 cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga
     1921 gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg
     1981 tttttgcgtg agccatgaga acgaaccatt gagatcatac ttactttgca tgtcactcaa
     2041 aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt
     2101 tcttagtccg ttatgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt
     2161 catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac
     2221 ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct
     2281 gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc
     2341 atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt
     2401 tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga
     2461 gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg
     2521 gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt
     2581 ggcatagttt gtccactgga aaatcccaaa gcctttaacc aaaggattcc tgatttccac
     2641 agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat tttccctact
     2701 gatgttcatc atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt
     2761 agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg
     2821 ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga
     2881 catacatctc aattggtcta ggtgatttta atcactatac caattgagat gggctagtca
     2941 atgataatta catgtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt
     3001 tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt
     3061 tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat
     3121 aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta
     3181 caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg
     3241 cttaagtagc accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca
     3301 tcaggcacct gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg
     3361 cagtgaatgg gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac
     3421 ccataataca agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat
     3481 gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac
     3541 cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg
     3601 gtatcatcaa caggcttacc cgtcttactg tccctagtgc ttggattctc accaataaaa
     3661 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     3721 ggatctatca acaggagtcc aagcgagctc gtaaacttgg tctgacagct ctagctccgg
     3781 caaaaaaacg ggcaaggtgt caccaccctg ccctttttct ttaaaaccga aaagattact
     3841 tcgcgtttgc cacctgacgt ctaagaaaag gaatattcag caatttgccc gtgc
//
