LOCUS       sequence-399194-        7038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  1/2 truncation of NORAD mRNA tagged with 4 copies of Riboglow 
            (variant A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7038)
  AUTHORS   Sarfraz N, Moscoso E, Oertel T, Lee HJ, Ranjit S, Braselmann E
  TITLE     Visualizing orthogonal RNAs simultaneously in live mammalian cells 
            by fluorescence lifetime imaging microscopy (FLIM).
  JOURNAL   Nat Commun. 2023 Feb 16;14(1):867. doi: 10.1038/s41467-023-36531-y.
  PUBMED    36797241
REFERENCE   2  (bases 1 to 7038)
  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: "2023-02-16"; volume: "14(1):867. doi"; pages: " 
            10.1038/s41467-023-36531-y"
COMMENT     Sequence Label: 1/2-NORAD-(A)4x
FEATURES             Location/Qualifiers
     source          1..7038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2853..2864
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3826..3947
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3863..3882)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3917..3936
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3954..4409)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4091..4112)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4303..4322
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4436..4540
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4542..4899
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4542..4733
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4565..4585)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4750..4885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4934..5728
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4934..5728
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4988..5007)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5598..5617
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5916..5935)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5960..6007
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6336..6924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6825..6844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcagtt
      601 ccggtccggc agagatcgcg gagagacgca gaacgcagcc cgctcctcca gggccctcca
      661 ggccctccgg ccccgggccg gcgggtgaac tggggggccc cgggacaggc cgagccctct
      721 gccctgcaga taacggaggc ctctgctgtg gctgcccact ggctgtgccc gcccactggc
      781 tgtgcccaga ccttgaagcc gcagcgaacc tctctttccc accccacctc ggtgactaat
      841 ggcggccgtg gcgtctccca gcccggaccc cgccggcacc cgggtctccc gacccaagcc
      901 tcgacgaaac ccccgcagag ccgccgggac gcagcgcctt tgggcggcgc tgggcgtggt
      961 gggccgggaa gtatggcggc agctcgaacg ccgcgcggcg gaggccatta aggcgtggac
     1021 ggcccgggaa ggcggcctag ggacgcaagc aggctcggcc gcctctttag gccacggagc
     1081 cgcgcagatc cggttcccgg gtgaccactc tgtcgccatt gggcgagacc tacctagtcc
     1141 tgacgacaac ggacaaaggc cttaaggggc ctggaaggtg agcgaagtcc cgaacgacga
     1201 cgggtggaac ggttagcggc catcgggcgg ttggtcttca ttctaccaga ctttgctgtc
     1261 ggaagagaga aatggtagaa tgacaggcca cgtttggccc gttggaaatg cccaccaccc
     1321 tctgggaaga tttactggcc gtttatggaa ggcctgtgta tataatatga aaaagctgct
     1381 ctcaactcca ccccaacctt ttaatagaaa acatttgtca catctagccc ttctagatgg
     1441 aaagaggttg ccgacgtatg ataaaataga gttagaaagt tacacatctt gtaaattctc
     1501 atttgtttaa aagaaatcat agaaaataca tgtcttctgg agatgacttt tggaaatgga
     1561 gttgttaaga cggcctctgg aagcgatacg tccacgtttg ttaagtgggt tagatgacat
     1621 ggagctggaa gacctgagaa ggaagagaag aaggttctat gctagactgg tcatatttag
     1681 aagacatttt catattctat ccattgtttt gtgtgcattt tattcctcac tactgtgtat
     1741 atagttgaca atgctaagct tttttgaaat gtctcttctt tttagatgtt ctgaagtgcc
     1801 tgatatgtta aaattagagg tagcaaaatc acattttgta aatacctttt tgttacaatt
     1861 cataggaaat atttttgggg gggaatggcc aaatcacctg ttgagtaata ctcattgtgt
     1921 ttgtgcagtg gttcagggga ggagagagga gggggaggtg cagagagctc tatgccatcc
     1981 tgtttacagc gaggcaagat gaatcattat gtctgtgcat tttgttttac ttatctgtgt
     2041 atatagtgta cataaaggac agacgagtcc taattgacaa catctagtct ttctggatgt
     2101 taaagaggtt gccagtgtat gacaaaagta gagttagtaa actaatatat tttgtacatt
     2161 ttgttttaca agtcctagga aagattgtct tctgaaaatt tgatgtcttc tgggttgatg
     2221 gagatgggaa gggttctagg ccagaatgtt cacatttgga agactctttc aaattataac
     2281 tgttgttaca tgtttgcagt ttattcaaga ctgctgtata catagtagac aaattaactc
     2341 cttacttgaa acatctagtc tatctagatg tttagaagtg cccgatgtat gttaaatgta
     2401 taggtagtaa aataccactt tgtaaatatc tttttgctaa aattcatagg aaatgctttt
     2461 ggaaattgaa ttgtgaagcc acctttgtga acagtatagt aatgtctata cttgttcaat
     2521 agtttagagg aggtaggagg gaagaaattg caaaaggtaa tattactagt gtgttcatac
     2581 ttggacattt tcagacacca tttttctata tgttttgtgc attttgtttt gctctgtata
     2641 tagtatatat aatggacaaa tagtcctaat ttttcaacat ctagtctcta gatgttaaag
     2701 aggttgccag tgtatgacaa aggagtaaaa ttagcatatt ttgtacactt tgtgttgaaa
     2761 ttcgtaggaa aacttgtctt ctgtaaagac ttttgcatag gaatttgttt gaccatctct
     2821 aagcattaca cgtgcctgta cttgtccact ggattgaagg cagagaagga agggaggagg
     2881 gaatgattca aggccaaaat ggccacattt agaagatacc tcagatgata accattgtta
     2941 tgtgtgtgca attttattta acagtgctgt gtatgtggtg gacaagttat atgaaatatc
     3001 tagtctttct agatatttgg aagtgcttga tgtatttaaa agtggtagta gaataacact
     3061 ttgtaaatag cttttaaaaa ctgatgggaa atgctgtttg gaagtggaat tgttgaacca
     3121 cctgggaggt gggagggaag aaattgcaaa tggtgttttg ccattgttta ttagaaaatt
     3181 tcagcttaat ccattgtgta tatgttacat gcatttcatt taactttgct atactgtata
     3241 gctagcattc tgcagtcgac ggtaccggcc taaaagcgta gtgggaaagt gacgtgaaat
     3301 tcgtccagat tacttgatac ggttatactc cgaatgccac ctaggccata caacgagcaa
     3361 ggagactcag gtaccggcct aaaagcgtag tgggaaagtg acgtgaaatt cgtccagatt
     3421 acttgatacg gttatactcc gaatgccacc taggccatac aacgagcaag gagactcagg
     3481 atccggccta aaagcgtagt gggaaagtga cgtgaaattc gtccagatta cttgatacgg
     3541 ttatactccg aatgccacct aggccataca acgagcaagg agactcagga tccggcctaa
     3601 aagcgtagtg ggaaagtgac gtgaaattcg tccagattac ttgatacggt tatactccga
     3661 atgccaccta ggccatacaa cgagcaagga gactcaggat ccaccggatc tagataactg
     3721 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3781 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3841 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3901 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     3961 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4021 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4081 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4141 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4201 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4261 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4321 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4381 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4441 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4501 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4561 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4621 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4681 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4741 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4801 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4861 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4921 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4981 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5041 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5101 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5161 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5221 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5281 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5341 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5401 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5461 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5521 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5581 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5641 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5701 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5761 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5821 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5881 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5941 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     6001 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     6061 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     6121 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     6181 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6241 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6301 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6361 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6421 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6481 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6541 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6601 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6661 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6721 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6781 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6841 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6901 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6961 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7021 cgtattaccg ccatgcat
//
