LOCUS       sequence-210865-        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        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"
     polyA_signal    2201..2322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2238..2257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2292..2311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2329..2784)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2466..2487)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2678..2697
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2811..2915
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2917..3274
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2917..3108
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2940..2960)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3125..3260
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3309..4103
                     /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             3309..4103
                     /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(3363..3382)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3973..3992
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4291..4310)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4335..4382
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4711..5299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5200..5219
                     /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 gctagcatgg
      601 atgatgatat cgccgcgctc gtcgtcgaca acggctccgg catgtgcaag gccggcttcg
      661 cgggcgacga tgccccccgg gccgtcttcc cctccatcgt ggggcgcccc aggcaccagg
      721 gcgtgatggt gggcatgggt cagaaggatt cctatgtggg cgacgaggcc cagagcaaga
      781 gaggcatcct caccctgaag taccccatcg agcacggcat cgtcaccaac tgggacgaca
      841 tggagaaaat ctggcaccac accttctaca atgagctgcg tgtggctccc gaggagcacc
      901 ccgtgctgct gaccgaggcc cccctgaacc ccaaggccaa ccgcgagaag atgacccaga
      961 tcatgtttga gaccttcaac accccagcca tgtacgttgc tatccaggct gtgctatccc
     1021 tgtacgcctc tggccgtacc actggcatcg tgatggactc cggtgacggg gtcacccaca
     1081 ctgtgcccat ctacgagggg tatgccctcc cccatgccat cctgcgtctg gacctggctg
     1141 gccgggacct gactgactac ctcatgaaga tcctcaccga gcgcggctac agcttcacca
     1201 ccacggccga gcgggaaatc gtgcgtgaca ttaaggagaa gctgtgctac gtcgccctgg
     1261 acttcgagca agagatggcc acggctgctt ccagctcctc cctggagaag agctacgagc
     1321 tgcctgacgg ccaggtcatc accattggca atgagcggtt ccgctgccct gaggcactct
     1381 tccagccttc cttcctgggc atggagtcct gtggcatcca cgaaactacc ttcaactcca
     1441 tcatgaagtg tgacgtggac atccgcaaag acctgtacgc caacacagtg ctgtctggcg
     1501 gcaccaccat gtaccctggc attgccgaca ggatgcagaa ggagatcact gccctggcac
     1561 ccagcacaat gaagatcaag atcattgctc ctcctgagcg caagtactcc gtgtggatcg
     1621 gcggctccat cctggcctcg ctgtccacct tccagcagat gtggatcagc aagcaggagt
     1681 atgacgagtc cggcccctcc atcgtccacc gcaaatgctt ctaggctagc attctgcagt
     1741 cgacggtacc ggcctaaaag cgtagtggga aagtgacgtg aaattcgtcc agattacttg
     1801 atacggttat actccgaatg ccacctaggc catacaacga gcaaggagac tcaggtaccg
     1861 gcctaaaagc gtagtgggaa agtgacgtga aattcgtcca gattacttga tacggttata
     1921 ctccgaatgc cacctaggcc atacaacgag caaggagact caggatccgg cctaaaagcg
     1981 tagtgggaaa gtgacgtgaa attcgtccag attacttgat acggttatac tccgaatgcc
     2041 acctaggcca tacaacgagc aaggagactc aggatccacc ggatctagat aactgatcat
     2101 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2161 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2281 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     2341 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2401 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2461 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2521 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2581 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2641 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2701 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2761 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2821 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2881 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     2941 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3001 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3061 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3121 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3181 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3241 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     3301 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3481 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     3841 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4081 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4141 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4201 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4261 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4321 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     4381 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4441 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4501 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     4561 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4621 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4681 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4741 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4801 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4861 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4921 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4981 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5041 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5101 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5161 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5221 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5281 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5341 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5401 taccgccatg cat
//
