LOCUS       sequence-403636-        6702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing luciferase driven by human ACSL4 promoter (bases -1 to 
            -1899 with respect to exon1 start site).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6702)
  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 6702)
  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-(-1899)
FEATURES             Location/Qualifiers
     source          1..6702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     254..275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    543..591
                     /note="synthetic polyadenylation signal"
     misc_feature    605..696
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     645..664
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2669..2678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2675..4327
                     /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(2741..2759)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4368..4489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4379..4398)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4433..4452
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4737..4754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4908..5496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4988..5007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5667..6458,6459..6527))
                     /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     6290..6309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6528..6632)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6659..6702,1..412)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
       61 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
      121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
      181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
      241 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
      301 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
      361 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg
      421 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      481 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc
      541 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
      601 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
      661 tccccagtgc aagtgcaggt gccagaacat ttctctatcg ataggtaccg agctcttacg
      721 cgtgctagcc cgggctcgag aacttcagta agctgaagac tctgttcaga ggaaattacc
      781 taattgaaaa tgtctgggct aagagaccga caataaagga aactggcaaa agtactcttt
      841 aattacaagt tctttttcat cttcatccag cacactgatt tatgtgctac actgggtttg
      901 tattatatct atattttaat tccacaacta gacactaact ttattgagaa cagcgatatt
      961 catttcttca atttcatttg tgttaaccca actgtaggtg atagtgctct gtacatcata
     1021 gttacccagt gaattacgta cctgacatat gattgcttat ttctggtatt aaaacaagat
     1081 ttcataaaat aaacatgaca attaaactcg ttgtttgaag tactgaactt tgtaggtcaa
     1141 ttttgaaaaa ggaagctaca tgactgtctt agcaatattt ataggtatat tatatgtaag
     1201 tgggtgggtt tcttggaaat tgaattggta atatggcttt agagtagtaa cacaaaaact
     1261 atagctctgt gctctcagct aaccactttt agtgtcacct gggctgctta ttaaaaattc
     1321 aaatttctta tccctaccca gatcagaatc aaaatacctg ggggttgtag ctccaggtac
     1381 ctacatttca acaagcaccc cagataattg tgatgcacac tgaagaacta ctctagcttg
     1441 taatctcagg tggtaaggca ttttatatat acatatatat atacacacac acaagcacaa
     1501 atggtaattc tatgggaaaa tggaaatcag caagtgaagt gaaatgtcaa ttaccatcag
     1561 gaacaggaag ctagttcacc attacgcatc agggaactgg aagaattctt gagatttatt
     1621 tttttgggtg atatattctt ggcagtttat caggcctcat ttgcaattat ggtctgtccc
     1681 ttttgaagcc ctcagatagt tcgctggcat tttaatttta atagcaattt cttgcatctg
     1741 ttggaacagc attcgtatga ccggcttata agcaatgatc agggctagta agatgtcaca
     1801 aagctgcggt gacttttcct tatcagttgg gggttttttt gccatgatac catcgtggaa
     1861 atgaaagaca acgtgaaaat cctgcgtcct tgtgatattt catcagtggg agtgagcggg
     1921 gatcttaacg cagtaagagg caagataaac ggatgagagg cattagttca ggcttacagg
     1981 agcaagatgc ttttagtcaa agtgttcaag atagctctct taaagaggag cagtgaaacc
     2041 caaaaatgag tgtcttttga gggcagctgg aagatgaccc gaggggaact tttatcctgg
     2101 ggtggagtcc tgcgaagcaa gccgcaggtg agggcgtggg ccaattctgc gcctcactgc
     2161 tgttaggcgc aaggacatcc cgaaacacac tcaacgacac cagcagcgag actgacagaa
     2221 gcggatcgcc agcgggagca acaacttgag gcgagccgtt agccgcctcg ccttcctcgc
     2281 agcgcccctc ccgcgccccg ggaagcccgt tcggctccgc cttttcgact gacgtcccgg
     2341 gcgagcgggg gcgggcgcgt gggtgggccg agccgtctgc agccagcgat tcggctggct
     2401 ctgccacacc accgcgcgcc cccgctccgc ccgcccctcc gggcgcgtct tttccgggct
     2461 cgcgctgagt cccgcctccg ccggctgtcc gggtgcgcgc gcgccgctgc ggctttttct
     2521 ctggcctccg ccgcgcgctc ctcctcgtcc cagcgctagc gggcacgcgg ttcctttttg
     2581 cgagctttcc gagtgccagg cgccggccgg ctgcgaagac gcggtgggcc gcccctccga
     2641 agcttggcat tccggtactg ttggtaaagc caccatggaa gacgccaaaa acataaagaa
     2701 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc
     2761 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt
     2821 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg
     2881 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt
     2941 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta
     3001 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc
     3061 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat
     3121 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac
     3181 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga
     3241 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc
     3301 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa
     3361 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat
     3421 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga
     3481 agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc
     3541 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt
     3601 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa
     3661 gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat
     3721 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt
     3781 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga
     3841 actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa
     3901 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga
     3961 cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt
     4021 ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt
     4081 cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca
     4141 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc
     4201 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa
     4261 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc
     4321 cgtgtaattc tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga
     4381 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4441 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     4501 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     4561 aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct
     4621 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     4681 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg
     4741 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4801 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4861 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4921 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4981 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5041 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5101 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5161 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5221 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5281 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5341 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5401 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5461 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5521 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5581 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5641 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5701 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5761 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5821 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5881 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5941 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6001 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6061 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6121 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6181 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6241 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6301 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6361 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6421 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6481 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6541 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6601 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6661 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg tt
//
