LOCUS       sequence-456877-        5409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Myc 5' untranslated region firefly luciferase reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5409)
  AUTHORS   Kovalski JR, Sarioglu G, Subramanyam V, Hernandez G, Rademaker G, 
            Oses-Prieto JA, Slota M, Mohan N, Yiakis K, Liu I, Wen KW, Kim GE, 
            Miglani S, Burlingame AL, Goodarzi H, Perera RM, Ruggero D
  TITLE     Functional screen identifies RBM42 as a mediator of oncogenic mRNA 
            translation specificity.
  JOURNAL   Nat Cell Biol. 2025 Feb 4. doi: 10.1038/s41556-024-01604-7.
  PUBMED    39905246
REFERENCE   2  (bases 1 to 5409)
  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 Cell 
            Biol. 2025 Feb 4. doi: 10.1038/s41556-024-01604-7."
COMMENT     Sequence Label: pGL3_5UTR_Myc_FLuc
FEATURES             Location/Qualifiers
     source          1..5409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             661..2313
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(727..745)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2372..2493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2383..2402)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2437..2456
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2741..2758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2912..3500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2992..3011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3671..4462,4463..4531))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4294..4313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4532..4636)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4663..5118
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4750..4769)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4960..4981
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5249..5297
                     /note="synthetic polyadenylation signal"
     misc_feature    5311..5402
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5351..5370
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
       61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
      181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      241 caaaaagctt aactcgctgt agtaattcca gcgagaggca gagggagcga gcgggcggcc
      301 ggctagggtg gaagagccgg gcgagcagag ctgcgctgcg ggcgtcctgg gaagggagat
      361 ccggagcgaa tagggggctt cgcctctggc ccagccctcc cgctgatccc ccagccagcg
      421 gtccgcaacc cttgccgcat ccacgaaact ttgcccatag cagcgggcgg gcactttgca
      481 ctggaactta caacacccga gcaaggacgc gactctcccg acgcggggag gctattctgc
      541 ccatttgggg acacttcccc gccgctgcca ggacccgctt ctctgaaagg ctctccttgc
      601 agctgcttag acgctggatt tttttcgggt agtggaaaac cagcagcctc ccgcgacgcc
      661 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga
      721 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
      781 gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc
      841 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
      901 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
      961 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt
     1021 tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
     1081 aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga
     1141 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
     1201 tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga
     1261 tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg
     1321 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
     1381 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
     1441 cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac
     1501 aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg
     1561 attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct
     1621 aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat
     1681 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     1741 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     1801 acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt
     1861 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     1921 ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct
     1981 ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa
     2041 caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt
     2101 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     2161 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac
     2221 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     2281 aaggccaaga agggcggaaa gatcgccgtg taattctagt ccgcggcata tgtctagagt
     2341 cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     2401 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2461 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2521 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     2581 ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     2641 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     2701 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     2761 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2821 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4501 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4561 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4621 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     4681 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4741 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4801 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4861 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4921 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4981 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5041 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5101 tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     5161 cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta
     5221 aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg
     5281 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     5341 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     5401 ctatcgata
//
