LOCUS       sequence-401791-        6456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RPS28 UTR luciferase assay plasmid (SCR UTR1, sites 1 and 2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6456)
  AUTHORS   Bryant CJ, McCool MA, Rosado Gonzalez GT, Abriola L, Surovtseva YV, 
            Baserga SJ
  TITLE     Discovery of novel microRNA mimic repressors of ribosome biogenesis.
  JOURNAL   Nucleic Acids Res. 2024 Feb 28;52(4):1988-2011. doi: 
            10.1093/nar/gkad1235.
  PUBMED    38197221
REFERENCE   2  (bases 1 to 6456)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2024-02-28- 28"; volume: "52"; issue: 
            "4"; pages: "1988-2011"
COMMENT     Sequence Label: psiCHECK2 RPS28 UTR1 SCR
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        61..252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          488..620
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     603..622
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        665..683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             693..1628
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1870..1918
                     /note="synthetic polyadenylation signal"
     promoter        1927..2678
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2708..2717
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2714..4366
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3673..3681
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4410..4531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4421..4440)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4475..4494
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4664..4768
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4769..4837,4838..5629)
                     /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     complement(4987..5006)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5800..6388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6289..6308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt
       61 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac
      481 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
      541 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
      601 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag
      661 tacttaatac gactcactat aggctagcca ccatggcttc caaggtgtac gaccccgagc
      721 aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc
      781 tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc gtgatttttc
      841 tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac atcgagcccg
      901 tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag agcgggaatg
      961 gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag ctgctgaacc
     1021 ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc tttcactact
     1081 cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga
     1141 tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc aagagcgaag
     1201 agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc ccaagcaaga
     1261 tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc aaggagaagg
     1321 gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt aagggaggca
     1381 agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc agcgacgatc
     1441 tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt gtcgagggag
     1501 ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc agccaggagg
     1561 acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg
     1621 agcagtaatt ctaggcgatc gctcgaggct tggctgctcg ctgggtcttg gatgtcgggt
     1681 tcgaccactt ggccgatggg aatggtctgt caccttattt gcccgttttt tgtccgccac
     1741 acgtaccttc ggtgaactat taaataaagc gtttgtgttt caagttaact caggttcttg
     1801 tctgggttat acgactaggg tttctccagg tttcttgagt ggctcccagg cggtgcggcc
     1861 gctggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgag
     1921 gatctaaatg agtcttcgga cctcgcgggg gccgcttaag cggtggttag ggtttgtctg
     1981 acgcgggggg agggggaagg aacgaaacac tctcattcgg aggcggctcg gggtttggtc
     2041 ttggtggcca cgggcacgca gaagagcgcc gcgatcctct taagcacccc cccgccctcc
     2101 gtggaggcgg gggtttggtc ggcgggtggt aactggcggg ccgctgactc gggcgggtcg
     2161 cgcgccccag agtgtgacct tttcggtctg ctcgcagacc cccgggcggc gccgccgcgg
     2221 cggcgacggg ctcgctgggt cctaggctcc atggggaccg tatacgtgga caggctctgg
     2281 agcatccgca cgactgcggt gatattaccg gagaccttct gcgggacgag ccgggtcacg
     2341 cggctgacgc ggagcgtccg ttgggcgaca aacaccagga cggggcacag gtacactatc
     2401 ttgtcacccg gaggcgcgag ggactgcagg agcttcaggg agtggcgcag ctgcttcatc
     2461 cccgtggccc gttgctcgcg tttgctggcg gtgtccccgg aagaaatata tttgcatgtc
     2521 tttagttcta tgatgacaca aaccccgccc agcgtcttgt cattggcgaa ttcgaacacg
     2581 cagatgcagt cggggcggcg cggtcccagg tccacttcgc atattaaggt gacgcgtgtg
     2641 gcctcgaaca ccgagcgacc ctgcagcgac ccgcttaaaa gcttggcatt ccggtactgt
     2701 tggtaaagcc accatggccg atgctaagaa cattaagaag ggccctgctc ccttctaccc
     2761 tctggaggat ggcaccgctg gcgagcagct gcacaaggcc atgaagaggt atgccctggt
     2821 gcctggcacc attgccttca ccgatgccca cattgaggtg gacatcacct atgccgagta
     2881 cttcgagatg tctgtgcgcc tggccgaggc catgaagagg tacggcctga acaccaacca
     2941 ccgcatcgtg gtgtgctctg agaactctct gcagttcttc atgccagtgc tgggcgccct
     3001 gttcatcgga gtggccgtgg cccctgctaa cgacatttac aacgagcgcg agctgctgaa
     3061 cagcatgggc atttctcagc ctaccgtggt gttcgtgtct aagaagggcc tgcagaagat
     3121 cctgaacgtg cagaagaagc tgcctatcat ccagaagatc atcatcatgg actctaagac
     3181 cgactaccag ggcttccaga gcatgtacac attcgtgaca tctcatctgc ctcctggctt
     3241 caacgagtac gacttcgtgc cagagtcttt cgacagggac aaaaccattg ccctgatcat
     3301 gaacagctct gggtctaccg gcctgcctaa gggcgtggcc ctgcctcatc gcaccgcctg
     3361 tgtgcgcttc tctcacgccc gcgaccctat tttcggcaac cagatcatcc ccgacaccgc
     3421 tattctgagc gtggtgccat tccaccacgg cttcggcatg ttcaccaccc tgggctacct
     3481 gatttgcggc tttcgggtgg tgctgatgta ccgcttcgag gaggagctgt tcctgcgcag
     3541 cctgcaagac tacaaaattc agtctgccct gctggtgcca accctgttca gcttcttcgc
     3601 taagagcacc ctgatcgaca agtacgacct gtctaacctg cacgagattg cctctggcgg
     3661 cgccccactg tctaaggagg tgggcgaagc cgtggccaag cgctttcatc tgccaggcat
     3721 ccgccagggc tacggcctga ccgagacaac cagcgccatt ctgattaccc cagagggcga
     3781 cgacaagcct ggcgccgtgg gcaaggtggt gccattcttc gaggccaagg tggtggacct
     3841 ggacaccggc aagaccctgg gagtgaacca gcgcggcgag ctgtgtgtgc gcggccctat
     3901 gattatgtcc ggctacgtga ataaccctga ggccacaaac gccctgatcg acaaggacgg
     3961 ctggctgcac tctggcgaca ttgcctactg ggacgaggac gagcacttct tcatcgtgga
     4021 ccgcctgaag tctctgatca agtacaaggg ctaccaggtg gccccagccg agctggagtc
     4081 tatcctgctg cagcacccta acattttcga cgccggagtg gccggcctgc ccgacgacga
     4141 tgccggcgag ctgcctgccg ccgtcgtcgt gctggaacac ggcaagacca tgaccgagaa
     4201 ggagatcgtg gactatgtgg ccagccaggt gacaaccgcc aagaagctgc gcggcggagt
     4261 ggtgttcgtg gacgaggtgc ccaagggcct gaccggcaag ctggacgccc gcaagatccg
     4321 cgagatcctg atcaaggcta agaaaggcgg caagatcgcc gtgtaataat tctagagtcg
     4381 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     4441 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4501 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4561 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     4621 taaaatcgat aaggatccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4681 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4741 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4801 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4861 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4921 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4981 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5041 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5101 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5161 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5221 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5281 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5341 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5401 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5461 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5521 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5581 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5641 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5701 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5761 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5821 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5881 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5941 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6001 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6061 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6121 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6181 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6241 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6301 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6361 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6421 ggccttttgc tggccttttg ctcacatggc tcgaca
//
