LOCUS       sequence-260255-        5829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of luciferase driven by eIF3B promoter region mutated at 
            the E-box binding motif 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5829)
  AUTHORS   Santasusagna S, Zhu S, Jawalagatti V, Carceles-Cordon M, Ertel A, 
            Garcia-Longarte S, Song WM, Fujiwara N, Li P, Mendizabal I, Petrylak
            DP, Kelly WK, Reddy EP, Wang L, Schiewer MJ, Lujambio A, Karnes J, 
            Knudsen KE, Cordon-Cardo C, Dong H, Huang H, Carracedo A, Hoshida Y,
            Rodriguez-Bravo V, Domingo-Domenech J
  TITLE     Master transcription factor reprograming unleashes selective 
            translation promoting castration resistance and immune evasion in 
            lethal prostate cancer.
  JOURNAL   Cancer Discov. 2023 Sep 7. doi: 10.1158/2159-8290.CD-23-0306.
  PUBMED    37676710
REFERENCE   2  (bases 1 to 5829)
  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: "Cancer 
            Discov. 2023 Sep 7. doi: 10.1158/2159-8290.CD-23-0306."
COMMENT     Sequence Label: pGL4-eIF3B MBE2 mutant
FEATURES             Location/Qualifiers
     source          1..5829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1681..1690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1687..3339
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    3383..3504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3394..3413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3448..3467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3752..3769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3923..4511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4003..4022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4711..5502,5503..5571))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5676..5724
                     /note="synthetic polyadenylation signal"
     misc_feature    5738..5829
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5778..5797
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcacagggtc tcgctatgtt gccctgactg
       61 gtcttgaact cttgggctca agtgatcctc ctgtcttggc ctcccaaagt actgggaaga
      121 gaggtgtgag ccaccgcgca gggcctagcc attttaatat gaagtttagt atacagttca
      181 gtggcatcaa gtacattcac catgttgtac catcaccacc gtcatctcca aaacgttctc
      241 atcctcccaa attgaaggtc cgtccccgat aaacactccc cactctccac cccagcctct
      301 gggtaccacc tgttttattt atttatttaa ttttcgtaga gatgagggct ggcgtgcggg
      361 gggtggtggt gctatgttgg caaggctggt ctccaactcc taagctcaag cgatcttccc
      421 acctcagcct cccaaagtgc tgggataaca ggcgtgatcc actgggttgt tttctaggat
      481 gattaagaca tcccccaacc agtggttgtc ctccactccg gcaaaaaaca agatagaagc
      541 aaaaaacacc caatgcccgt cccgtgagca cccgcgcaga gcgaaccgct ccggtccctc
      601 cggagtcagc ctcgagctcc acgcggcgag gacccgcccc cgcccccacg tgacccgcgc
      661 tgtcccgcgc tgaaggcggc ggcctcggat ggactgaacc actgtgaaag cccgaatggg
      721 gggaaacagg cagtgacggc cgctagggaa ggactggggg taagcgatta aagaaacttc
      781 catacatttt tgtatcgcgc caggcagtcg tccttctagt cgtgtgggcc ctctctggct
      841 cgccccgaat tccttttcta ttttgaagta gtctgtcctt ttgctcagtc agtaggcgag
      901 tccaagtgac agacacggac aggggggact ttagtaacgt aaactaaacg gtttccctag
      961 cgaaaggctc agacgccctg agcaatcgcc tgttgagtct ccactcttcc agggtattaa
     1021 ctgttgagtt agaggacatt ttcctagcat tccgggtact aaaatcattt cttgtccccg
     1081 ccttcagctc cacaaatagt tcagtcccgc cccgcagcgt ccgacgggaa aggaaagggg
     1141 gaggacgcca tgttggacgg aaccatctgc tgccacacat ggggtggtca atgctcccat
     1201 gagtttactg atgcacagaa acaaccttgc tgaatggaat taagcacttt cttgtcacaa
     1261 atatatcagt ttgatgattt tcgggcgcgg tcttgtggat gcgagtcctc tcagggtcgt
     1321 tcgtgaggcc ccagggcgtg ggtgcgcccc ccgccccgcg gccttggtgc ggcctccccg
     1381 tcgcacgcac atggctgggc tgtagccgtc gcggcgcgcg gtgcggcctg ggagagtcgg
     1441 aagcgcggcg gccgcggagc cctgcgagta ggcagcgttg ggcccatgca ggacgcggag
     1501 aacgtggcgg tgcccgaggc ggccgaggag cgcgccgagc ccggccagca gcagccggcc
     1561 gccgagccgc cgccagccga ggggctgctg cggcccgcgg ggcccggcgc tccggaggcc
     1621 gcggggaccg agagatctgg cctcggcggc caagcttggc aatccggtac tgttggtaaa
     1681 gccaccatgg aagatgccaa aaacattaag aagggcccag cgccattcta cccactcgaa
     1741 gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc
     1801 accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag
     1861 atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc
     1921 gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc
     1981 ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg
     2041 ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac
     2101 gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac
     2161 cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag
     2221 tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt
     2281 agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga
     2341 ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc
     2401 agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc
     2461 ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa
     2521 gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc
     2581 actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg
     2641 ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag
     2701 ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag
     2761 cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc
     2821 ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg
     2881 agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg
     2941 cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg
     3001 aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg
     3061 ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc
     3121 gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc
     3181 gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc
     3241 gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt
     3301 ctcattaagg ccaagaaggg cggcaagatc gccgtgtaat aattctagag tcggggcggc
     3361 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3421 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3481 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3541 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     3601 gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     3661 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     3721 aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4321 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4681 gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc ttgatcagtg aggcaccgat
     4741 ctcagcgatc tgcctatttc gttcgtccat agtggcctga ctccccgtcg tgtagatcac
     4801 tacgattcgt gagggcttac catcaggccc cagcgcagca atgatgccgc gagagccgcg
     4861 ttcaccggcc cccgatttgt cagcaatgaa ccagccagca gggagggccg agcgaagaag
     4921 tggtcctgct actttgtccg cctccatcca gtctatgagc tgctgtcgtg atgctagagt
     4981 aagaagttcg ccagtgagta gtttccgaag agttgtggcc attgctactg gcatcgtggt
     5041 atcacgctcg tcgttcggta tggcttcgtt caactctggt tcccagcggt caagccgggt
     5101 cacatgatca cccatattat gaagaaatgc agtcagctcc ttagggcctc cgatcgttgt
     5161 cagaagtaag ttggccgcgg tgttgtcgct catggtaatg gcagcactac acaattctct
     5221 taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc gagtactcaa ccaagtcgtt
     5281 ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg gcgtctatac gggacaacac
     5341 cgcgccacat agcagtactt tgaaagtgct catcatcggg aatcgttctt cggggcggaa
     5401 agactcaagg atcttgccgc tattgagatc cagttcgata tagcccactc ttgcacccag
     5461 ttgatcttca gcatctttta ctttcaccag cgtttcgggg tgtgcaaaaa caggcaagca
     5521 aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt tggatgctca tactcgtcct
     5581 ttttcaatat tattgaagca tttatcaggg ttactagtac gtctctcaag gataagtaag
     5641 taatattaag gtacgggagg tattggacag gccgcaataa aatatcttta ttttcattac
     5701 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     5761 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     5821 catttctct
//
