LOCUS       sequence-214332-        5413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ACTB mRNA tagged with 3 copies of Riboglow (variant A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5413)
  AUTHORS   Braselmann E, Wierzba AJ, Polaski JT, Chrominski M, Holmes ZE, Hung 
            ST, Batan D, Wheeler JR, Parker R, Jimenez R, Gryko D, Batey RT, 
            Palmer AE
  TITLE     A multicolor riboswitch-based platform for imaging of RNA in live 
            mammalian cells.
  JOURNAL   Nat Chem Biol. 2018 Jul 30. pii: 10.1038/s41589-018-0103-7. doi: 
            10.1038/s41589-018-0103-7.
  PUBMED    30061719
REFERENCE   2  (bases 1 to 5413)
  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 Chem 
            Biol. 2018 Jul 30. pii: 10.1038/s41589-018-0103-7. doi: 
            10.1038/s41589-018-0103-7."
COMMENT     Sequence Label: ACTB-(A)3x
FEATURES             Location/Qualifiers
     source          1..5413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2196..2317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2233..2252)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2287..2306
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2324..2779)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2461..2482)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2673..2692
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2806..2910
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2912..3269
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2912..3103
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2935..2955)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3120..3255
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3304..4098
                     /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             3304..4098
                     /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(3358..3377)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3968..3987
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4286..4305)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4330..4377
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4706..5294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5195..5214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag catggatgat
      601 gatatcgccg cgctcgtcgt cgacaacggc tccggcatgt gcaaggccgg cttcgcgggc
      661 gacgatgccc cccgggccgt cttcccctcc atcgtggggc gccccaggca ccagggcgtg
      721 atggtgggca tgggtcagaa ggattcctat gtgggcgacg aggcccagag caagagaggc
      781 atcctcaccc tgaagtaccc catcgagcac ggcatcgtca ccaactggga cgacatggag
      841 aaaatctggc accacacctt ctacaatgag ctgcgtgtgg ctcccgagga gcaccccgtg
      901 ctgctgaccg aggcccccct gaaccccaag gccaaccgcg agaagatgac ccagatcatg
      961 tttgagacct tcaacacccc agccatgtac gttgctatcc aggctgtgct atccctgtac
     1021 gcctctggcc gtaccactgg catcgtgatg gactccggtg acggggtcac ccacactgtg
     1081 cccatctacg aggggtatgc cctcccccat gccatcctgc gtctggacct ggctggccgg
     1141 gacctgactg actacctcat gaagatcctc accgagcgcg gctacagctt caccaccacg
     1201 gccgagcggg aaatcgtgcg tgacattaag gagaagctgt gctacgtcgc cctggacttc
     1261 gagcaagaga tggccacggc tgcttccagc tcctccctgg agaagagcta cgagctgcct
     1321 gacggccagg tcatcaccat tggcaatgag cggttccgct gccctgaggc actcttccag
     1381 ccttccttcc tgggcatgga gtcctgtggc atccacgaaa ctaccttcaa ctccatcatg
     1441 aagtgtgacg tggacatccg caaagacctg tacgccaaca cagtgctgtc tggcggcacc
     1501 accatgtacc ctggcattgc cgacaggatg cagaaggaga tcactgccct ggcacccagc
     1561 acaatgaaga tcaagatcat tgctcctcct gagcgcaagt actccgtgtg gatcggcggc
     1621 tccatcctgg cctcgctgtc caccttccag cagatgtgga tcagcaagca ggagtatgac
     1681 gagtccggcc cctccatcgt ccaccgcaaa tgcttctagg ctagcattct gcagtcgacg
     1741 gtaccggcct aaaagcgtag tgggaaagtg acgtgaaatt cgtccagatt acttgatacg
     1801 gttatactcc gaatgccacc taggccatac aacgagcaag gagactcagg taccggccta
     1861 aaagcgtagt gggaaagtga cgtgaaattc gtccagatta cttgatacgg ttatactccg
     1921 aatgccacct aggccataca acgagcaagg agactcagga tccggcctaa aagcgtagtg
     1981 ggaaagtgac gtgaaattcg tccagattac ttgatacggt tatactccga atgccaccta
     2041 ggccatacaa cgagcaagga gactcaggat ccaccggatc tagataactg atcataatca
     2101 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2161 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2221 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2281 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     2341 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2401 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2461 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2521 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2581 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2641 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2701 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2761 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2821 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2881 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     2941 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3001 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3061 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3121 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3181 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3241 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3301 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3361 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3421 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3481 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3541 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3601 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3661 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3721 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3781 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3841 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3901 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3961 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4021 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4081 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4141 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4201 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4261 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4321 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4381 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4441 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4501 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4561 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4621 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4681 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4741 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4801 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4861 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4921 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4981 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5041 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5101 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5161 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5221 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5281 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5341 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5401 ccatgcatta gtt
//
