LOCUS       sequence-450180-        4673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MET 5'UTR (uORF2 mutated) upstream FLuc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4673)
  AUTHORS   Cerqua M, Foiani M, Boccaccio C, Comoglio PM, Altintas DM
  TITLE     The integrated stress response drives MET oncogene overexpression in
            cancers.
  JOURNAL   EMBO J. 2025 Jan 7. doi: 10.1038/s44318-024-00338-4.
  PUBMED    39774381
REFERENCE   2  (bases 1 to 4673)
  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: "EMBO J. 
            2025 Jan 7. doi: 10.1038/s44318-024-00338-4."
COMMENT     Sequence Label: pGL4.23[uORF2/luc2/minP]
FEATURES             Location/Qualifiers
     source          1..4673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        77..108
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             531..2183
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    2227..2348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2238..2257)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2292..2311
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2596..2613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2767..3355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2847..2866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3555..4346,4347..4415))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4520..4568
                     /note="synthetic polyadenylation signal"
     misc_feature    4582..4673
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4622..4641
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaagatc tggcctcggc
       61 ggccaagctt agacactaga gggtatataa tggaagctcg acttccagct tggcaatccg
      121 gtactgttgg taaaagacac gtgctggggc gggcaggcga gcgcctcagt ctggtcgcct
      181 ggcggtgcct ccggccccaa cgcgcccggg ccgccgcggg ccgcgcgcgc cgatgcccgg
      241 ctgagtcact ggcagggcag cgcgcgtgtg ggaaggggcg gagggagtgc ggccggcggg
      301 cgggcggggc gctgggctca gccgggccgc aggtgacccg gaggccctcg ccgcccgcgg
      361 cgccccgagc gctttgtgag cagttccgga gccgagtgga gggcgcgagc cagatgcggg
      421 gcgacagctg acttgctgag aggaggcggg gaggcgcgga gcgcgcgtgt ggtccttgcg
      481 ccgctgactt ctccactggt tcctgggcac cgaaagataa acctctcata atggaagatg
      541 ccaaaaacat taagaagggc ccagcgccat tctacccact cgaagacggg accgccggcg
      601 agcagctgca caaagccatg aagcgctacg ccctggtgcc cggcaccatc gcctttaccg
      661 acgcacatat cgaggtggac attacctacg ccgagtactt cgagatgagc gttcggctgg
      721 cagaagctat gaagcgctat gggctgaata caaaccatcg gatcgtggtg tgcagcgaga
      781 atagcttgca gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc
      841 cagctaacga catctacaac gagcgcgagc tgctgaacag catgggcatc agccagccca
      901 ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa aagaagctac
      961 cgatcataca aaagatcatc atcatggata gcaagaccga ctaccagggc ttccaaagca
     1021 tgtacacctt cgtgacttcc catttgccac ccggcttcaa cgagtacgac ttcgtgcccg
     1081 agagcttcga ccgggacaaa accatcgccc tgatcatgaa cagtagtggc agtaccggat
     1141 tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg
     1201 accccatctt cggcaaccag atcatccccg acaccgctat cctcagcgtg gtgccatttc
     1261 accacggctt cggcatgttc accacgctgg gctacttgat ctgcggcttt cgggtcgtgc
     1321 tcatgtaccg cttcgaggag gagctattct tgcgcagctt gcaagactat aagattcaat
     1381 ctgccctgct ggtgcccaca ctatttagct tcttcgctaa gagcactctc atcgacaagt
     1441 acgacctaag caacttgcac gagatcgcca gcggcggggc gccgctcagc aaggaggtag
     1501 gtgaggccgt ggccaaacgc ttccacctac caggcatccg ccagggctac ggcctgacag
     1561 aaacaaccag cgccattctg atcacccccg aaggggacga caagcctggc gcagtaggca
     1621 aggtggtgcc cttcttcgag gctaaggtgg tggacttgga caccggtaag acactgggtg
     1681 tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat catgagcggc tacgttaaca
     1741 accccgaggc tacaaacgct ctcatcgaca aggacggctg gctgcacagc ggcgacatcg
     1801 cctactggga cgaggacgag cacttcttca tcgtggaccg gctgaagagc ctgatcaaat
     1861 acaagggcta ccaggtagcc ccagccgaac tggagagcat cctgctgcaa caccccaaca
     1921 tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg cccgccgcag
     1981 tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga gatcgtggac tatgtggcca
     2041 gccaggttac aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta
     2101 aaggactgac cggcaagttg gacgcccgca agatccgcga gattctcatt aaggccaaga
     2161 agggcggcaa gatcgccgtg taataattct agagtcgggg cggccggccg cttcgagcag
     2221 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2281 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2341 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2401 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga
     2461 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     2521 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     2581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagcggcc gcaaatgcta
     3541 aaccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc gatctgccta
     3601 tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat tcgtgagggc
     3661 ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc ggcccccgat
     3721 ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc tgctactttg
     3781 tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag ttcgccagtg
     3841 agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg ctcgtcgttc
     3901 ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg atcacccata
     3961 ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag taagttggcc
     4021 gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt catgccatcc
     4081 gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga gtagtgtata
     4141 cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc acatagcagt
     4201 actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc aaggatcttg
     4261 ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc ttcagcatct
     4321 tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc cgcaaagaag
     4381 ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca atattattga
     4441 agcatttatc agggttacta gtacgtctct caaggataag taagtaatat taaggtacgg
     4501 gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     4561 tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa
     4621 actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tct
//
