LOCUS       sequence-214329-        5195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ACTB mRNA tagged with 1 copy of Riboglow (variant A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5195)
  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 5195)
  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)1x
FEATURES             Location/Qualifiers
     source          1..5195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1981..2102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2018..2037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2072..2091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2109..2564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2246..2267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2458..2477
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2591..2695
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2697..3054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2697..2888
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2720..2740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2905..3040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3089..3883
                     /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             3089..3883
                     /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(3143..3162)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3753..3772
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4071..4090)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4115..4162
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4491..5079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4980..4999
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcatggat
      601 gatgatatcg ccgcgctcgt cgtcgacaac ggctccggca tgtgcaaggc cggcttcgcg
      661 ggcgacgatg ccccccgggc cgtcttcccc tccatcgtgg ggcgccccag gcaccagggc
      721 gtgatggtgg gcatgggtca gaaggattcc tatgtgggcg acgaggccca gagcaagaga
      781 ggcatcctca ccctgaagta ccccatcgag cacggcatcg tcaccaactg ggacgacatg
      841 gagaaaatct ggcaccacac cttctacaat gagctgcgtg tggctcccga ggagcacccc
      901 gtgctgctga ccgaggcccc cctgaacccc aaggccaacc gcgagaagat gacccagatc
      961 atgtttgaga ccttcaacac cccagccatg tacgttgcta tccaggctgt gctatccctg
     1021 tacgcctctg gccgtaccac tggcatcgtg atggactccg gtgacggggt cacccacact
     1081 gtgcccatct acgaggggta tgccctcccc catgccatcc tgcgtctgga cctggctggc
     1141 cgggacctga ctgactacct catgaagatc ctcaccgagc gcggctacag cttcaccacc
     1201 acggccgagc gggaaatcgt gcgtgacatt aaggagaagc tgtgctacgt cgccctggac
     1261 ttcgagcaag agatggccac ggctgcttcc agctcctccc tggagaagag ctacgagctg
     1321 cctgacggcc aggtcatcac cattggcaat gagcggttcc gctgccctga ggcactcttc
     1381 cagccttcct tcctgggcat ggagtcctgt ggcatccacg aaactacctt caactccatc
     1441 atgaagtgtg acgtggacat ccgcaaagac ctgtacgcca acacagtgct gtctggcggc
     1501 accaccatgt accctggcat tgccgacagg atgcagaagg agatcactgc cctggcaccc
     1561 agcacaatga agatcaagat cattgctcct cctgagcgca agtactccgt gtggatcggc
     1621 ggctccatcc tggcctcgct gtccaccttc cagcagatgt ggatcagcaa gcaggagtat
     1681 gacgagtccg gcccctccat cgtccaccgc aaatgcttct aggctagcat tctgcagtcg
     1741 acggtaccgg cctaaaagcg tagtgggaaa gtgacgtgaa attcgtccag attacttgat
     1801 acggttatac tccgaatgcc acctaggcca tacaacgagc aaggagactc aggatccacc
     1861 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1921 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1981 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2041 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2101 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2161 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2221 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2281 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2341 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2401 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2461 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2521 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     2581 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2641 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     2701 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2761 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2821 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2881 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2941 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3001 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3061 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3121 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3181 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3241 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3301 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3361 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3421 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3481 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3541 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3601 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3661 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3721 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3781 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3841 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3901 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3961 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4021 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4081 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4141 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4201 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4261 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4321 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4381 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4441 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4501 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4561 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4621 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4681 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4741 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4801 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4861 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4921 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4981 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5041 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5101 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5161 gattctgtgg ataaccgtat taccgccatg catta
//
