LOCUS       sequence-275162-        5339 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 5339)
  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 5339)
  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 500bp promoter
FEATURES             Location/Qualifiers
     source          1..5339
                     /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      702..711
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             708..2360
                     /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(774..792)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2401..2522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2412..2431)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2466..2485
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2770..2787)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2941..3529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3021..3040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3700..4491,4492..4560))
                     /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     4323..4342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4561..4665)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4692..5147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4779..4798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4989..5010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5278..5326
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgtg
      121 ctagccctct ctagtcccgc ctccctaacc tgattggttt attcaaacaa accccggcca
      181 actcagccgt tcataggtgg atataaaagg caagctacga ttggttcttc tggacggaga
      241 cggtgagagc gagtcaggga ttggctggtc tgcttcgggc gggctaaagg aaggttcaag
      301 tggagctctc ctaaccgacg cgcgtctgtg gagaagcggc ttggtcgggg gtggtctcgt
      361 ggggtcctgc ctgtttagtc gctttcaggg ttcttgagcc ccttcacgac cgtcaccatg
      421 gaagtgtcac cattgcaggt acggctcgcg ggggggactg gcggtggagc ctccgcgcgg
      481 cccgggcatc tctctggccg cccgtgacgg gtgaagctct ggggctgcgg tcaggccggc
      541 gaccggcttg ggagcccata ttctccattc ccggttccgg ggtgatcgtg gagaggcgga
      601 agccccttct ggtgctagta gtgaagaatt cgggtatata atggatccgg tatcgagatc
      661 tgcgatctaa gtaagcttgg cattccggta ctgttggtaa agccaccatg gaagacgcca
      721 aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc gctggagagc
      781 aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg
      841 cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt cggttggcag
      901 aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact
      961 ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg
     1021 cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg cagcctaccg
     1081 tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa
     1141 tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt
     1201 acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt gtgccagagt
     1261 ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct actggtctgc
     1321 ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat gccagagatc
     1381 ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc
     1441 acggttttgg aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa
     1501 tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag attcaaagtg
     1561 cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt gacaaatacg
     1621 atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag gaagtcgggg
     1681 aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg ctcactgaga
     1741 ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag
     1801 ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta
     1861 atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat gtaaacaatc
     1921 cggaagcgac caacgccttg attgacaagg atggatggct acattctgga gacatagctt
     1981 actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg attaagtaca
     2041 aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac cccaacatct
     2101 tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg
     2161 ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc
     2221 aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag
     2281 gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg
     2341 gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg gccgcttcga gcagacatga
     2401 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2461 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2521 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2581 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg
     2641 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     2701 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     2761 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2821 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2881 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2941 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3061 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3121 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3181 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3241 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3301 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3361 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3421 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3481 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3541 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3601 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3661 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3721 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3781 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3841 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3901 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3961 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4021 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4081 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4141 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4201 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4261 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4321 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4381 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4441 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4501 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4561 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4621 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4681 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4741 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4801 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4861 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4921 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4981 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5041 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5101 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt
     5161 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc
     5221 caagctacca tgataagtaa gtaatattaa ggtacgggag gtacttggag cggccgcaat
     5281 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtact
//
