LOCUS       sequence-401790-        6456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RPS28 UTR luciferase assay plasmid (WT 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 WT
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1871..1919
                     /note="synthetic polyadenylation signal"
     promoter        1928..2679
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2709..2718
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2715..4367
                     /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             3674..3682
                     /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    4411..4532
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4422..4441)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4476..4495
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4665..4769
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4770..4838,4839..5630)
                     /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(4988..5007)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5801..6389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6290..6309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgaggc ttggctgctc gctgggtctt ggatgtcggg
     1681 ttcgaccact tggccgatgg gaatggtctg tcacaatctg ctcctttttt ttgtccgcca
     1741 cacgtaactg agatgctcct ttaaataaag cgtttgtgtt tcaagttaac tcaggttctt
     1801 gtctgggtta tacgactagg gtttctccag gtttcttgag tggctcccag gcggtgcggc
     1861 cgctggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     1921 ggatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct
     1981 gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt
     2041 cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc
     2101 cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc
     2161 gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg
     2221 gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg
     2281 gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac
     2341 gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat
     2401 cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat
     2461 ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt
     2521 ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac
     2581 gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt
     2641 ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttggcat tccggtactg
     2701 ttggtaaagc caccatggcc gatgctaaga acattaagaa gggccctgct cccttctacc
     2761 ctctggagga tggcaccgct ggcgagcagc tgcacaaggc catgaagagg tatgccctgg
     2821 tgcctggcac cattgccttc accgatgccc acattgaggt ggacatcacc tatgccgagt
     2881 acttcgagat gtctgtgcgc ctggccgagg ccatgaagag gtacggcctg aacaccaacc
     2941 accgcatcgt ggtgtgctct gagaactctc tgcagttctt catgccagtg ctgggcgccc
     3001 tgttcatcgg agtggccgtg gcccctgcta acgacattta caacgagcgc gagctgctga
     3061 acagcatggg catttctcag cctaccgtgg tgttcgtgtc taagaagggc ctgcagaaga
     3121 tcctgaacgt gcagaagaag ctgcctatca tccagaagat catcatcatg gactctaaga
     3181 ccgactacca gggcttccag agcatgtaca cattcgtgac atctcatctg cctcctggct
     3241 tcaacgagta cgacttcgtg ccagagtctt tcgacaggga caaaaccatt gccctgatca
     3301 tgaacagctc tgggtctacc ggcctgccta agggcgtggc cctgcctcat cgcaccgcct
     3361 gtgtgcgctt ctctcacgcc cgcgacccta ttttcggcaa ccagatcatc cccgacaccg
     3421 ctattctgag cgtggtgcca ttccaccacg gcttcggcat gttcaccacc ctgggctacc
     3481 tgatttgcgg ctttcgggtg gtgctgatgt accgcttcga ggaggagctg ttcctgcgca
     3541 gcctgcaaga ctacaaaatt cagtctgccc tgctggtgcc aaccctgttc agcttcttcg
     3601 ctaagagcac cctgatcgac aagtacgacc tgtctaacct gcacgagatt gcctctggcg
     3661 gcgccccact gtctaaggag gtgggcgaag ccgtggccaa gcgctttcat ctgccaggca
     3721 tccgccaggg ctacggcctg accgagacaa ccagcgccat tctgattacc ccagagggcg
     3781 acgacaagcc tggcgccgtg ggcaaggtgg tgccattctt cgaggccaag gtggtggacc
     3841 tggacaccgg caagaccctg ggagtgaacc agcgcggcga gctgtgtgtg cgcggcccta
     3901 tgattatgtc cggctacgtg aataaccctg aggccacaaa cgccctgatc gacaaggacg
     3961 gctggctgca ctctggcgac attgcctact gggacgagga cgagcacttc ttcatcgtgg
     4021 accgcctgaa gtctctgatc aagtacaagg gctaccaggt ggccccagcc gagctggagt
     4081 ctatcctgct gcagcaccct aacattttcg acgccggagt ggccggcctg cccgacgacg
     4141 atgccggcga gctgcctgcc gccgtcgtcg tgctggaaca cggcaagacc atgaccgaga
     4201 aggagatcgt ggactatgtg gccagccagg tgacaaccgc caagaagctg cgcggcggag
     4261 tggtgttcgt ggacgaggtg cccaagggcc tgaccggcaa gctggacgcc cgcaagatcc
     4321 gcgagatcct gatcaaggct aagaaaggcg gcaagatcgc cgtgtaataa ttctagagtc
     4381 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     4441 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4501 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     4561 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     4621 gtaaaatcga taaggatcca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4681 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4741 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4801 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4861 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4921 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4981 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5041 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5101 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5161 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5221 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5281 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5341 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5401 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5461 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5521 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5581 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5641 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5701 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5761 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5821 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5881 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5941 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6001 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6061 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6121 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6181 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6241 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6301 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6361 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6421 tggccttttg ctggcctttt gctcacatgg ctcgac
//
