LOCUS       sequence-217237-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the S(-31)-M30 apta-FRET RNA origami structure..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  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 5570)
  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(-31)-M30_dT7term
FEATURES             Location/Qualifiers
     source          1..5570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      384..431
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             446..463
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(516..534)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      530..577
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      614..1069
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(701..720)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     911..932
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1161..1976)
                     /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(1161..1976)
                     /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     1884..1903
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1968..1987
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2098..2686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2587..2606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2840..2857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2872..3011
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2937..2956)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3097..3119
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3113..3304)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4113..5195)
                     /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     5157..5176
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5196..5273)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5479..5498
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg agatcgagcg acttccgact tcggtcggga gtcgccgtgc
       61 ggtaaggacg ggtccctagc taattcgtta gttaggttga gtagagtgtg agaccgtacg
      121 ggctggtgaa gcctccacgc cagcctcggt ctcccgcagt aggatcggac tgaaggaggc
      181 acggtcccag ccgaagtgtc ttgcttcggc aaggcacttt ggctgctaga ctggctggtt
      241 cgttcgatac gcagcgaagg gacggtgcgg agaggagagc tgcgtgtcga acggaccagc
      301 tagtttagga ttctattgcg aattccctag cataaccccg cggggcctct tcgggggtct
      361 cgcggggttt tttgctgaaa gacctagcat aaccccttgg ggcctctaaa cgggtcttga
      421 ggggtttttt gctgaaagac tcgagcacca ccaccaccac cactgagatc cggctgctaa
      481 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
      541 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
      601 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
      661 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      721 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      781 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      841 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      901 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      961 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1021 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     1081 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1141 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     1201 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     1261 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     1321 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     1381 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     1441 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     1501 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     1561 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     1621 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     1681 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     1741 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     1801 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     1861 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     1921 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca
     1981 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta
     2041 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2101 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2161 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2221 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2281 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2341 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2401 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2461 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2521 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2581 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2641 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2701 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2761 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2821 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2881 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     2941 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3001 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3061 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3121 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3181 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     3241 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     3301 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     3361 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     3421 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     3481 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     3541 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     3601 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     3661 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     3721 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca
     3781 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     3841 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     3901 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     3961 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     4021 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     4081 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4141 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4201 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     4261 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     4321 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     4381 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     4441 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     4501 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     4561 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     4621 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     4681 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     4741 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     4801 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     4861 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     4921 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     4981 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     5041 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     5101 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     5161 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     5221 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     5281 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     5341 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     5401 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     5461 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     5521 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctttc
//
