LOCUS       sequence-378531-        4418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter for duplicated TERT promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4418)
  AUTHORS   Barger CJ, Suwala AK, Soczek KM, Wang AS, Kim MY, Hong C, Doudna JA,
            Chang SM, Phillips JJ, Solomon DA, Costello JF
  TITLE     Conserved features of TERT promoter duplications reveal an 
            activation mechanism that mimics hotspot mutations in cancer.
  JOURNAL   Nat Commun. 2022 Sep 16;13(1):5430. doi: 10.1038/s41467-022-33099-x.
  PUBMED    36114166
REFERENCE   2  (bases 1 to 4418)
  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 
            Commun."; date: "2022-09-16"; volume: "13(1):5430. doi"; pages: " 
            10.1038/s41467-022-33099-x"
COMMENT     Sequence Label: pGL4.0-TERT_-100_-79
FEATURES             Location/Qualifiers
     source          1..4418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1672..1793
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1683..1702)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1737..1756
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2041..2058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2212..2800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2292..2311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3000..3791,3792..3860))
                     /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    3965..4013
                     /note="synthetic polyadenylation signal"
     misc_feature    4027..4118
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4067..4086
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      4388..4397
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4394..4418,1..1628)
                     /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"
ORIGIN
        1 agggcccagc gccattctac ccactcgaag acgggaccgc cggcgagcag ctgcacaaag
       61 ccatgaagcg ctacgccctg gtgcccggca ccatcgcctt taccgacgca catatcgagg
      121 tggacattac ctacgccgag tacttcgaga tgagcgttcg gctggcagaa gctatgaagc
      181 gctatgggct gaatacaaac catcggatcg tggtgtgcag cgagaatagc ttgcagttct
      241 tcatgcccgt gttgggtgcc ctgttcatcg gtgtggctgt ggccccagct aacgacatct
      301 acaacgagcg cgagctgctg aacagcatgg gcatcagcca gcccaccgtc gtattcgtga
      361 gcaagaaagg gctgcaaaag atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga
      421 tcatcatcat ggatagcaag accgactacc agggcttcca aagcatgtac accttcgtga
      481 cttcccattt gccacccggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg
      541 acaaaaccat cgccctgatc atgaacagta gtggcagtac cggattgccc aagggcgtag
      601 ccctaccgca ccgcaccgct tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca
      661 accagatcat ccccgacacc gctatcctca gcgtggtgcc atttcaccac ggcttcggca
      721 tgttcaccac gctgggctac ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg
      781 aggaggagct attcttgcgc agcttgcaag actataagat tcaatctgcc ctgctggtgc
      841 ccacactatt tagcttcttc gctaagagca ctctcatcga caagtacgac ctaagcaact
      901 tgcacgagat cgccagcggc ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca
      961 aacgcttcca cctaccaggc atccgccagg gctacggcct gacagaaaca accagcgcca
     1021 ttctgatcac ccccgaaggg gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct
     1081 tcgaggctaa ggtggtggac ttggacaccg gtaagacact gggtgtgaac cagcgcggcg
     1141 agctgtgcgt ccgtggcccc atgatcatga gcggctacgt taacaacccc gaggctacaa
     1201 acgctctcat cgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg
     1261 acgagcactt cttcatcgtg gaccggctga agagcctgat caaatacaag ggctaccagg
     1321 tagccccagc cgaactggag agcatcctgc tgcaacaccc caacatcttc gacgccgggg
     1381 tcgccggcct gcccgacgac gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac
     1441 acggtaaaac catgaccgag aaggagatcg tggactatgt ggccagccag gttacaaccg
     1501 ccaagaagct gcgcggtggt gttgtgttcg tggacgaggt gcctaaagga ctgaccggca
     1561 agttggacgc ccgcaagatc cgcgagattc tcattaaggc caagaagggc ggcaagatcg
     1621 ccgtgtaata attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     1681 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     1741 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     1801 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     1861 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga
     1921 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     1981 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     2041 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2101 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2401 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2641 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2941 aagtatatat gagtaaactt ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt
     3001 taccagtgct tgatcagtga ggcaccgatc tcagcgatct gcctatttcg ttcgtccata
     3061 gtggcctgac tccccgtcgt gtagatcact acgattcgtg agggcttacc atcaggcccc
     3121 agcgcagcaa tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc agcaatgaac
     3181 cagccagcag ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag
     3241 tctatgagct gctgtcgtga tgctagagta agaagttcgc cagtgagtag tttccgaaga
     3301 gttgtggcca ttgctactgg catcgtggta tcacgctcgt cgttcggtat ggcttcgttc
     3361 aactctggtt cccagcggtc aagccgggtc acatgatcac ccatattatg aagaaatgca
     3421 gtcagctcct tagggcctcc gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc
     3481 atggtaatgg cagcactaca caattctctt accgtcatgc catccgtaag atgcttttcc
     3541 gtgaccggcg agtactcaac caagtcgttt tgtgagtagt gtatacggcg accaagctgc
     3601 tcttgcccgg cgtctatacg ggacaacacc gcgccacata gcagtacttt gaaagtgctc
     3661 atcatcggga atcgttcttc ggggcggaaa gactcaagga tcttgccgct attgagatcc
     3721 agttcgatat agcccactct tgcacccagt tgatcttcag catcttttac tttcaccagc
     3781 gtttcggggt gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca
     3841 cgaaaatgtt ggatgctcat actcgtcctt tttcaatatt attgaagcat ttatcagggt
     3901 tactagtacg tctctcaagg ataagtaagt aatattaagg tacgggaggt attggacagg
     3961 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
     4021 agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc
     4081 tgtccccagt gcaagtgcag gtgccagaac atttctctgg cctaactggc cggtacctga
     4141 gctcgctagc ctcgagtcct gccccttcac cttccagctc cgcctcctcc gcgcggaccc
     4201 cgccccgtcc cgacccctcc cgggtccccg gcccagcccc ctccgggccc tcccagcccc
     4261 tccccttcct ttccgcggcc ccgccccttc ctttccgcgg ccccgccctc tcctcgcggc
     4321 gcgagtttca ggcagcgctg gatctggcct cggcggccaa gcttggcaat ccggtactgt
     4381 tggtaaagcc accatggaag atgccaaaaa cattaaga
//
