LOCUS       sequence-266885-        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"
     regulatory      1103..1112
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1109..2761
                     /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(1175..1193)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2802..2923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2813..2832)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2867..2886
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3171..3188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3342..3930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3422..3441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4101..4892,4893..4961))
                     /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     4724..4743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4962..5066)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5093..5548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5180..5199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5390..5411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5679..5727
                     /note="synthetic polyadenylation signal"
     misc_feature    5741..5832
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5781..5800
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagccatt cccctcgcta acaacgtcag aacagaggac
       61 agtttttaga tttcagggat cttaaataga ttggcagttc ctggagaata aacatccttt
      121 gcttttctcc tgcacacttt tgcctcaggc caccccttcc cgcttccaaa gcccatctct
      181 tccaagcttt ccgcacgaga aaacaagtga gcccttctca ttggccagat tccctgtcaa
      241 tctctccgct atgacgccga gtggtgcctt ttgaagcctc tctagtcccg cctccctaac
      301 ctgattggtt tattcaaaca aaccccggcc aactcagccg ttcataggtg gatataaaag
      361 gcaagctacg attggttctt ctggacggag acggtgagag cgagtcaggg attggctggt
      421 ctgcttcggg cgggctaaag gaaggttcaa gtggagctct cctaaccgac gcgcgtctgt
      481 ggagaagcgg cttggtcggg ggtggtctcg tggggtcctg cctgtttagt cgctttcagg
      541 gttcttgagc cccttcacga ccgtcaccat ggaagtgtca ccattgcagg tacggctcgc
      601 gggggggact ggcggtggag cctccgcgcg gcccgggcat ctctctggcc gcccgtgacg
      661 ggtgaagctc tggggctgcg gtcaggccgg cgaccggctt gggagcccat attctccatt
      721 cccggttccg gggtgatcgt ggagaggcgg aagccccttc tggtgctagt agtgaagtat
      781 gacccggctt ccagggtgtc gtgcgttgtt gccctgtttc taggggcagg agtccgttgg
      841 tcccgtagtg gatccatgtt acagcggccc gggtgcgacg ttattgagtc gcgcgtacag
      901 atgctttccc ctcctgcccg ctgcttgaaa acggtcttga atgtccccga tcttggaaag
      961 ggcagagcct agcacagtgt ttctctggag cagctggcca gcttacaaaa gcgccccctt
     1021 cctgaggaat tcgggtatat aatggatccg gtatcgagat ctgcgatcta agtaagcttg
     1081 gcattccggt actgttggta aagccaccat ggaagacgcc aaaaacataa agaaaggccc
     1141 ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata aggctatgaa
     1201 gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtggacat
     1261 cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg
     1321 gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc
     1381 ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga
     1441 acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa
     1501 ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa aaattattat
     1561 catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca
     1621 tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata gggacaagac
     1681 aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg tcgctctgcc
     1741 tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg gcaatcaaat
     1801 cattccggat actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac
     1861 tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga
     1921 gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg tgccaaccct
     1981 attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga
     2041 aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt
     2101 ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag ctattctgat
     2161 tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc
     2221 gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg
     2281 tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt
     2341 gattgacaag gatggatggc tacattctgg agacatagct tactgggacg aagacgaaca
     2401 cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc aggtggctcc
     2461 cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg
     2521 tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa
     2581 gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa
     2641 gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga
     2701 cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta
     2761 attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2821 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2881 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2941 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     3001 ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc
     3061 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     3121 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     3181 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3241 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3301 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3361 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3421 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3481 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3541 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3601 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3661 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3781 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4081 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4201 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4261 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4321 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4381 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     5101 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5161 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5221 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5281 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5341 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5401 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5461 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5521 tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg
     5581 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta
     5641 agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat
     5701 tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa
     5761 acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca
     5821 gaacatttct ctatcgata
//
