LOCUS       sequence-375337-        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"
     primer_bind     22..41
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      343..352
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             349..2001
                     /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    2045..2166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2056..2075)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2110..2129
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2414..2431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2585..3173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2665..2684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3373..4164,4165..4233))
                     /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    4338..4386
                     /note="synthetic polyadenylation signal"
     misc_feature    join(4400..4418,1..73)
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc
       61 agaacatttc tctggcctaa ctggccggta cctgagctcg ctagcctcga gtcctgcccc
      121 ttcaccttcc agctccgcct cctccgcgcg gaccccgccc cgtcccgacc cctcccgggt
      181 ccccggccca gccccctccg ggccctccca gcccctcccc ttcctttccg cggccccgcc
      241 ccttcctttc cgcggccccg ccctctcctc gcggcgcgag tttcaggcag cgctggatct
      301 ggcctcggcg gccaagcttg gcaatccggt actgttggta aagccaccat ggaagatgcc
      361 aaaaacatta agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag
      421 cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac
      481 gcacatatcg aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca
      541 gaagctatga agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat
      601 agcttgcagt tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca
      661 gctaacgaca tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc
      721 gtcgtattcg tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg
      781 atcatacaaa agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg
      841 tacaccttcg tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag
      901 agcttcgacc gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg
      961 cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac
     1021 cccatcttcg gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac
     1081 cacggcttcg gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc
     1141 atgtaccgct tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct
     1201 gccctgctgg tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac
     1261 gacctaagca acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt
     1321 gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa
     1381 acaaccagcg ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag
     1441 gtggtgccct tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg
     1501 aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac
     1561 cccgaggcta caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc
     1621 tactgggacg aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac
     1681 aagggctacc aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc
     1741 ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc
     1801 gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc
     1861 caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa
     1921 ggactgaccg gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag
     1981 ggcggcaaga tcgccgtgta ataattctag agtcggggcg gccggccgct tcgagcagac
     2041 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2101 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2161 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2221 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     2281 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     2341 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     2401 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2461 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3001 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3121 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc aaatgctaaa
     3361 ccactgcagt ggttaccagt gcttgatcag tgaggcaccg atctcagcga tctgcctatt
     3421 tcgttcgtcc atagtggcct gactccccgt cgtgtagatc actacgattc gtgagggctt
     3481 accatcaggc cccagcgcag caatgatgcc gcgagagccg cgttcaccgg cccccgattt
     3541 gtcagcaatg aaccagccag cagggagggc cgagcgaaga agtggtcctg ctactttgtc
     3601 cgcctccatc cagtctatga gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag
     3661 tagtttccga agagttgtgg ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg
     3721 tatggcttcg ttcaactctg gttcccagcg gtcaagccgg gtcacatgat cacccatatt
     3781 atgaagaaat gcagtcagct ccttagggcc tccgatcgtt gtcagaagta agttggccgc
     3841 ggtgttgtcg ctcatggtaa tggcagcact acacaattct cttaccgtca tgccatccgt
     3901 aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg
     3961 gcgaccaagc tgctcttgcc cggcgtctat acgggacaac accgcgccac atagcagtac
     4021 tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc
     4081 gctattgaga tccagttcga tatagcccac tcttgcaccc agttgatctt cagcatcttt
     4141 tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg
     4201 aatgagtgcg acacgaaaat gttggatgct catactcgtc ctttttcaat attattgaag
     4261 catttatcag ggttactagt acgtctctca aggataagta agtaatatta aggtacggga
     4321 ggtattggac aggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     4381 tgtgtgaatc gatagtacta acatacgctc tccatcaa
//
