LOCUS       sequence-396603-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing luciferase driven by human ACSL4 promoter (bases -1 to 
            -254 with respect to exon1 start site).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  AUTHORS   Wang ME, Chen J, Lu Y, Bawcom AR, Wu J, Ou J, Asara JM, Armstrong 
            AJ, Wang Q, Li L, Wang Y, Huang J, Chen M
  TITLE     RB1-deficient prostate tumor growth and metastasis are vulnerable to
            ferroptosis induction via the E2F/ACSL4 axis.
  JOURNAL   J Clin Invest. 2023 Mar 16:e166647. doi: 10.1172/JCI166647.
  PUBMED    36928314
REFERENCE   2  (bases 1 to 5057)
  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: "J Clin 
            Invest. 2023 Mar 16:e166647. doi: 10.1172/JCI166647."
COMMENT     Sequence Label: pGL3-ACSL4-(-254)
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    398..446
                     /note="synthetic polyadenylation signal"
     misc_feature    460..551
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     500..519
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      879..888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             885..2537
                     /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(951..969)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2578..2699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2589..2608)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2643..2662
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2947..2964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3118..3706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3198..3217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3877..4668,4669..4737))
                     /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     4500..4519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4738..4842)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(4869..5057,1..267)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4956..4975)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
       61 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      121 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
      181 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      241 ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg
      301 aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa
      361 gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt
      421 acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa
      481 caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag
      541 aacatttctc tatcgatagg taccgagctc ttacgcgtgc tagcccgggc tcgaggcgat
      601 tcggctggct ctgccacacc accgcgcgcc cccgctccgc ccgcccctcc gggcgcgtct
      661 tttccgggct cgcgctgagt cccgcctccg ccggctgtcc gggtgcgcgc gcgccgctgc
      721 ggctttttct ctggcctccg ccgcgcgctc ctcctcgtcc cagcgctagc gggcacgcgg
      781 ttcctttttg cgagctttcc gagtgccagg cgccggccgg ctgcgaagac gcggtgggcc
      841 gcccctccga agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa
      901 acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac
      961 tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac
     1021 atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag
     1081 ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc
     1141 ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga
     1201 acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg
     1261 tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca
     1321 tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca
     1381 cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct
     1441 tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta
     1501 aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta
     1561 tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg
     1621 gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt
     1681 atagatttga agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc
     1741 tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt
     1801 tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag
     1861 cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta
     1921 catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg
     1981 ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc
     2041 aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg
     2101 aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact
     2161 gggacgaaga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag
     2221 gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg
     2281 acgcaggtgt cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg
     2341 ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag
     2401 taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc
     2461 ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg
     2521 gaaagatcgc cgtgtaattc tagagtcggg gcggccggcc gcttcgagca gacatgataa
     2581 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2641 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2701 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     2761 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgatgccc
     2821 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     2881 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
     2941 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3001 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3061 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3121 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3181 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3241 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3301 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3361 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3421 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3481 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3541 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3601 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3661 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3721 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3781 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3841 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3901 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3961 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4021 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4081 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4141 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4201 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4261 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4321 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4381 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4441 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4501 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4561 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4621 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4681 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4741 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4801 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4861 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4921 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4981 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5041 ccgatttagt gctttac
//
