LOCUS       sequence-217238-        5502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the S*5-M5 apta-FRET RNA origami structure..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5502)
  AUTHORS   Jepsen MDE, Sparvath SM, Nielsen TB, Langvad AH, Grossi G, Gothelf 
            KV, Andersen ES
  TITLE     Development of a genetically encodable FRET system using fluorescent
            RNA aptamers.
  JOURNAL   Nat Commun. 2018 Jan 2;9(1):18. doi: 10.1038/s41467-017-02435-x.
  PUBMED    29295996
REFERENCE   2  (bases 1 to 5502)
  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: "2018-01-2"; volume: "9(1):18. doi"; pages: " 
            10.1038/s41467-017-02435-x"
COMMENT     Sequence Label: pET28c_S*5-M5_dT7term
FEATURES             Location/Qualifiers
     source          1..5502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      297..344
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             359..376
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(429..447)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      443..490
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      527..982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(614..633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     824..845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1074..1889)
                     /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"
     CDS             complement(1074..1889)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /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     1797..1816
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1881..1900
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2011..2599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2500..2519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2753..2770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2785..2924
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2850..2869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3010..3032
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3026..3217)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4026..5108)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5070..5089
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5109..5186)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5392..5411
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(5484..5502,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5484..5502
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gagagaccgc cctcgctgtc ttcggacagt gagggggctg tcctgcttcg gcagggcagc
       61 cgattgtgaa gcctccacgc aatccggttt ctcccacgcg ggttgcgggt tgaaggaggc
      121 acgacccctg ggttgagtag agtgtgaggg gctacggctt cggccgtggc ccaaggacgg
      181 gtccccagcc atcgaaggga cggtgcggag aggagagatg ggtaacctgc gtgaattccc
      241 tagcataacc ccgcggggcc tcttcggggg tctcgcgggg ttttttgctg aaagacctag
      301 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa gactcgagca
      361 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
      421 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
      481 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
      541 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      601 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      661 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      721 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
      781 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      841 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
      901 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
      961 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     1081 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1141 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1201 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     1261 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1321 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     1381 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     1441 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     1501 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     1561 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     1621 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     1681 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     1741 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     1801 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     1861 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     1921 gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1981 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2041 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2101 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2161 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2221 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2281 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2341 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2401 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2461 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2521 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2581 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2641 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2701 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2761 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2821 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2881 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     2941 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3001 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3061 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3121 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     3181 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     3241 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     3301 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     3361 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     3421 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     3481 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     3541 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     3601 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     3661 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     3721 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     3781 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     3841 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     3901 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     3961 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4021 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4081 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4141 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     4201 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     4261 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     4321 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     4381 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     4441 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     4501 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     4561 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     4621 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     4681 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     4741 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     4801 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     4861 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     4921 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     4981 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5041 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5101 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     5161 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     5221 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     5281 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     5341 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     5401 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     5461 ggcgtagagg atcgagatct ttctaatacg actcactata gg
//
