LOCUS       sequence-377362-        3878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription template plasmid for RPLP1 5'UTR followed by Renilla 
            luciferase open reading frame and polyA sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3878)
  AUTHORS   Calviello L, Venkataramanan S, Rogowski KJ, Wyler E, Wilkins K, 
            Tejura M, Thai B, Krol J, Filipowicz W, Landthaler M, Floor SN
  TITLE     DDX3 depletion represses translation of mRNAs with complex 5' UTRs.
  JOURNAL   Nucleic Acids Res. 2021 May 21;49(9):5336-5350. doi: 
            10.1093/nar/gkab287.
  PUBMED    33905506
REFERENCE   2  (bases 1 to 3878)
  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: "2021-05-21- 21"; volume: "49"; issue: 
            "9"; pages: "5336-5350"
COMMENT     Sequence Label: pUC57-RPLP15UTR-hRluc-A60
FEATURES             Location/Qualifiers
     source          1..3878
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..60)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     160..179
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     373..395
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     387..404
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     388..404
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        417..435
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             601..1536
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     primer_bind     complement(1666..1682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1666..1682)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1679..1701)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1690..1706
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1714..1720,1721..1738,1739..1744))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1759..1780
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1897..1914)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2068..2656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2148..2167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2827..3618,3619..3687))
                     /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     3450..3469
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3688..3792)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3860..3878
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
       61 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      121 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      181 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      241 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      301 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      361 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctctaat
      421 acgactcact ataggtggac acataagagg ctgcgtatag gcgcgagagc ccctttcctc
      481 agctgccgcc aaggtgctcg gtccttccga ggaagctaag gctgcgttgg ggtgaggccc
      541 tcacttcatc cggcgactag caccgcgtcc ggcagcgcca gccctacact cgcccgcgcc
      601 atggcttcca aggtgtacga ccccgagcaa cgcaaacgca tgatcactgg gcctcagtgg
      661 tgggctcgct gcaagcaaat gaacgtgctg gactccttca tcaactacta tgattccgag
      721 aagcacgccg agaacgccgt gatttttctg catggtaacg ctgcctccag ctacctgtgg
      781 aggcacgtcg tgcctcacat cgagcccgtg gctagatgca tcatccctga tctgatcgga
      841 atgggtaagt ccggcaagag cgggaatggc tcatatcgcc tcctggatca ctacaagtac
      901 ctcaccgctt ggttcgagct gctgaacctt ccaaagaaaa tcatctttgt gggccacgac
      961 tggggggctt gtctggcctt tcactactcc tacgagcacc aagacaagat caaggccatc
     1021 gtccatgctg agagtgtcgt ggacgtgatc gagtcctggg acgagtggcc tgacatcgag
     1081 gaggatatcg ccctgatcaa gagcgaagag ggcgagaaaa tggtgcttga gaataacttc
     1141 ttcgtcgaga ccatgctccc aagcaagatc atgcggaaac tggagcctga ggagttcgct
     1201 gcctacctgg agccattcaa ggagaagggc gaggttagac ggcctaccct ctcctggcct
     1261 cgcgagatcc ctctcgttaa gggaggcaag cccgacgtcg tccagattgt ccgcaactac
     1321 aacgcctacc ttcgggccag cgacgatctg cctaagatgt tcatcgagtc cgaccctggg
     1381 ttcttttcca acgctattgt cgagggagct aagaagttcc ctaacaccga gttcgtgaag
     1441 gtgaagggcc tccacttcag ccaggaggac gctccagatg aaatgggtaa gtacatcaag
     1501 agcttcgtgg agcgcgtgct gaagaacgag cagtaattct aggcgatcgc tcgagcccgg
     1561 gaattcgttt aaacctagag cggccgctgg ccgcctgcag aaaaaaaaaa aatgcatctg
     1621 gtttcctggt ctgtctcctc tcggtttaag cttggcgtaa tcatggtcat agctgtttcc
     1681 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     1741 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1801 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1861 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1921 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1981 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2041 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2101 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2161 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2221 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2281 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2341 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2401 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2461 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2521 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2581 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2641 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2701 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2761 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2821 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2881 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2941 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3001 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3061 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3121 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3181 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3241 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3301 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3361 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3421 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3481 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3541 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3601 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3661 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3721 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3781 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     3841 catgacatta acctataaaa ataggcgtat cacgaggc
//
