LOCUS       sequence-266881-        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"
     regulatory      603..612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             609..2261
                     /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(675..693)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2302..2423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2313..2332)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2367..2386
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2671..2688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2842..3430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2922..2941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3601..4392,4393..4461))
                     /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     4224..4243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4462..4566)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4593..5048
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4680..4699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4890..4911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5179..5227
                     /note="synthetic polyadenylation signal"
     misc_feature    5241..5332
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5281..5300
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagccctc tctagtcccg cctccctaac ctgattggtt
       61 tattcaaaca aaccccggcc aactcagccg ttcataggtg gatataaaag gcaagctacg
      121 attggttctt ctggacggag acggtgagag cgagtcaggg attggctggt ctgcttcggg
      181 cgggctaaag gaaggttcaa gtggagctct cctaaccgac gcgcgtctgt ggagaagcgg
      241 cttggtcggg ggtggtctcg tggggtcctg cctgtttagt cgctttcagg gttcttgagc
      301 cccttcacga ccgtcaccat ggaagtgtca ccattgcagg tacggctcgc gggggggact
      361 ggcggtggag cctccgcgcg gcccgggcat ctctctggcc gcccgtgacg ggtgaagctc
      421 tggggctgcg gtcaggccgg cgaccggctt gggagcccat attctccatt cccggttccg
      481 gggtgatcgt ggagaggcgg aagccccttc tggtgctagt agtgaagaat tcgggtatat
      541 aatggatccg gtatcgagat ctgcgatcta agtaagcttg gcattccggt actgttggta
      601 aagccaccat ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg
      661 aagatggaac cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg
      721 gaacaattgc ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg
      781 aaatgtccgt tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa
      841 tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta
      901 tcggagttgc agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta
      961 tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga
     1021 acgtgcaaaa aaagctccca atcatccaaa aaattattat catggattct aaaacggatt
     1081 accagggatt tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg
     1141 aatacgattt tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact
     1201 cctctggatc tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga
     1261 gattctcgca tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt
     1321 taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat
     1381 gtggatttcg agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc
     1441 aggattacaa gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa
     1501 gcactctgat tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc
     1561 ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc
     1621 aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag ggggatgata
     1681 aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata
     1741 ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta
     1801 tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc
     1861 tacattctgg agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc
     1921 tgaagtctct gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct
     1981 tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg
     2041 gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga
     2101 tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt
     2161 ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga
     2221 tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta attctagagt cggggcggcc
     2281 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2341 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2401 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2461 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     2521 ataaggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     2581 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     2641 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3241 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3901 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4201 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4561 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     4621 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4681 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4741 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4801 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4861 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4921 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4981 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5041 tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5101 ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga
     5161 ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     5221 ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa
     5281 ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgata
//
