LOCUS       sequence-214330-        5304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ACTB mRNA tagged with 2 copies of Riboglow (variant A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5304)
  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 5304)
  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)2x
FEATURES             Location/Qualifiers
     source          1..5304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     515..535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2088..2209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2125..2144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2179..2198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2216..2671)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2353..2374)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2565..2584
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2698..2802
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2804..3161
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2804..2995
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2827..2847)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3012..3147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3196..3990
                     /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             3196..3990
                     /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(3250..3269)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3860..3879
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4178..4197)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4222..4269
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4598..5186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5087..5106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatggatga
      601 tgatatcgcc gcgctcgtcg tcgacaacgg ctccggcatg tgcaaggccg gcttcgcggg
      661 cgacgatgcc ccccgggccg tcttcccctc catcgtgggg cgccccaggc accagggcgt
      721 gatggtgggc atgggtcaga aggattccta tgtgggcgac gaggcccaga gcaagagagg
      781 catcctcacc ctgaagtacc ccatcgagca cggcatcgtc accaactggg acgacatgga
      841 gaaaatctgg caccacacct tctacaatga gctgcgtgtg gctcccgagg agcaccccgt
      901 gctgctgacc gaggcccccc tgaaccccaa ggccaaccgc gagaagatga cccagatcat
      961 gtttgagacc ttcaacaccc cagccatgta cgttgctatc caggctgtgc tatccctgta
     1021 cgcctctggc cgtaccactg gcatcgtgat ggactccggt gacggggtca cccacactgt
     1081 gcccatctac gaggggtatg ccctccccca tgccatcctg cgtctggacc tggctggccg
     1141 ggacctgact gactacctca tgaagatcct caccgagcgc ggctacagct tcaccaccac
     1201 ggccgagcgg gaaatcgtgc gtgacattaa ggagaagctg tgctacgtcg ccctggactt
     1261 cgagcaagag atggccacgg ctgcttccag ctcctccctg gagaagagct acgagctgcc
     1321 tgacggccag gtcatcacca ttggcaatga gcggttccgc tgccctgagg cactcttcca
     1381 gccttccttc ctgggcatgg agtcctgtgg catccacgaa actaccttca actccatcat
     1441 gaagtgtgac gtggacatcc gcaaagacct gtacgccaac acagtgctgt ctggcggcac
     1501 caccatgtac cctggcattg ccgacaggat gcagaaggag atcactgccc tggcacccag
     1561 cacaatgaag atcaagatca ttgctcctcc tgagcgcaag tactccgtgt ggatcggcgg
     1621 ctccatcctg gcctcgctgt ccaccttcca gcagatgtgg atcagcaagc aggagtatga
     1681 cgagtccggc ccctccatcg tccaccgcaa atgcttctag gctagcattc tgcagtcgac
     1741 ggtaccggcc taaaagcgta gtgggaaagt gacgtgaaat tcgtccagat tacttgatac
     1801 ggttatactc cgaatgccac ctaggccata caacgagcaa ggagactcag gtaccggcct
     1861 aaaagcgtag tgggaaagtg acgtgaaatt cgtccagatt acttgatacg gttatactcc
     1921 gaatgccacc taggccatac aacgagcaag gagactcagg atccaccgga tctagataac
     1981 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2041 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2101 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2161 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     2221 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2281 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2341 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2401 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2461 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2521 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2581 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2641 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2701 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2761 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2821 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2881 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3001 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3061 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3121 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3181 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3241 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3301 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3361 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3421 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3481 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3541 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3601 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3661 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3721 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3781 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3841 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3901 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3961 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4021 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4081 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4141 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4201 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4261 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4321 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4381 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4441 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4501 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4561 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4621 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4681 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4741 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4801 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4861 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4921 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4981 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5041 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5101 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5161 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5221 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5281 accgtattac cgccatgcat tagt
//
