LOCUS       sequence-275163-        5839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter plasmid driven by TOP2A promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5839)
  AUTHORS   Kwan SY, Sheel A, Song CQ, Zhang XO, Jiang T, Dang H, Cao Y, Ozata 
            DM, Mou H, Yin H, Weng Z, Wang XW, Xue W
  TITLE     Depletion of TRRAP induces p53-independent senescence in liver 
            cancer by downregulating mitotic genes.
  JOURNAL   Hepatology. 2019 Jun 12. doi: 10.1002/hep.30807.
  PUBMED    31188495
REFERENCE   2  (bases 1 to 5839)
  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: "Hepatology.
            2019 Jun 12. doi: 10.1002/hep.30807."
COMMENT     Sequence Label: pBV-luc TOP2A 1kb promoter
FEATURES             Location/Qualifiers
     source          1..5839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      1202..1211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1208..2860
                     /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(1274..1292)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2901..3022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2912..2931)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2966..2985
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3270..3287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3441..4029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3521..3540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4200..4991,4992..5060))
                     /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     4823..4842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5061..5165)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5192..5647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5279..5298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5489..5510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5778..5826
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtg
      121 ctagccattc ccctcgctaa caacgtcaga acagaggaca gtttttagat ttcagggatc
      181 ttaaatagat tggcagttcc tggagaataa acatcctttg cttttctcct gcacactttt
      241 gcctcaggcc accccttccc gcttccaaag cccatctctt ccaagctttc cgcacgagaa
      301 aacaagtgag cccttctcat tggccagatt ccctgtcaat ctctccgcta tgacgccgag
      361 tggtgccttt tgaagcctct ctagtcccgc ctccctaacc tgattggttt attcaaacaa
      421 accccggcca actcagccgt tcataggtgg atataaaagg caagctacga ttggttcttc
      481 tggacggaga cggtgagagc gagtcaggga ttggctggtc tgcttcgggc gggctaaagg
      541 aaggttcaag tggagctctc ctaaccgacg cgcgtctgtg gagaagcggc ttggtcgggg
      601 gtggtctcgt ggggtcctgc ctgtttagtc gctttcaggg ttcttgagcc ccttcacgac
      661 cgtcaccatg gaagtgtcac cattgcaggt acggctcgcg ggggggactg gcggtggagc
      721 ctccgcgcgg cccgggcatc tctctggccg cccgtgacgg gtgaagctct ggggctgcgg
      781 tcaggccggc gaccggcttg ggagcccata ttctccattc ccggttccgg ggtgatcgtg
      841 gagaggcgga agccccttct ggtgctagta gtgaagtatg acccggcttc cagggtgtcg
      901 tgcgttgttg ccctgtttct aggggcagga gtccgttggt cccgtagtgg atccatgtta
      961 cagcggcccg ggtgcgacgt tattgagtcg cgcgtacaga tgctttcccc tcctgcccgc
     1021 tgcttgaaaa cggtcttgaa tgtccccgat cttggaaagg gcagagccta gcacagtgtt
     1081 tctctggagc agctggccag cttacaaaag cgcccccttc ctgaggaatt cgggtatata
     1141 atggatccgg tatcgagatc tgcgatctaa gtaagcttgg cattccggta ctgttggtaa
     1201 agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga
     1261 agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg
     1321 aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga
     1381 aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat
     1441 cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat
     1501 cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat
     1561 gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa
     1621 cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta
     1681 ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga
     1741 atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc
     1801 ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag
     1861 attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt
     1921 aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat atttgatatg
     1981 tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca
     2041 ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag
     2101 cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc
     2161 cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca
     2221 aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa
     2281 accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac
     2341 cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat
     2401 gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct
     2461 acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct
     2521 gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt
     2581 gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg
     2641 tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat
     2701 cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt
     2761 tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat
     2821 cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg
     2881 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2941 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3001 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3061 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3121 taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     3181 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     3241 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5161 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     5221 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5281 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5341 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5401 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5461 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5521 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5581 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     5641 acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5701 tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag
     5761 gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     5821 tgtgtgaatc gatagtact
//
