LOCUS       sequence-223058-        4271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector to produce humanized Renilla luciferase with 5BoxB
            sequences in the 3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4271)
  AUTHORS   Pillai RS, Artus CG, Filipowicz W
  TITLE     Tethering of human Ago proteins to mRNA mimics the miRNA-mediated 
            repression of protein synthesis.
  JOURNAL   RNA. 2004 Oct . 10(10):1518-25.
  PUBMED    15337849
REFERENCE   2  (bases 1 to 4271)
  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: "RNA. 2004 
            Oct . 10(10):1518-25."
COMMENT     Sequence Label: phRL TK 5BoxB sp 36
FEATURES             Location/Qualifiers
     source          1..4271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        664..1415
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1485..1617
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1600..1619
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1662..1680
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1684..1693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1690..2625
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     protein_bind    2662..2680
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    2706..2724
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    2730..2748
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    2789..2807
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    2813..2831
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     polyA_signal    2882..3003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2893..2912)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2947..2966
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3136..3240
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3241..3309,3310..4101)
                     /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(3459..3478)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
      661 ctaaatgagt cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg
      721 cggggggagg gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg
      781 gtggccacgg gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg
      841 gaggcggggg tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc
      901 gccccagagt gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg
      961 cgacgggctc gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc
     1021 atccgcacga ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg
     1081 ctgacgcgga gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg
     1141 tcacccggag gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc
     1201 gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt
     1261 agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag
     1321 atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc
     1381 tcgaacaccg agcgaccctg cagcgacccg cttaaaagct tgattcttct gacacaacag
     1441 tctcgaactt aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta
     1501 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
     1561 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
     1621 tccactccca gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg
     1681 ctagccacca tggcttccaa ggtgtacgac cccgagcaac gcaaacgcat gatcactggg
     1741 cctcagtggt gggctcgctg caagcaaatg aacgtgctgg actccttcat caactactat
     1801 gattccgaga agcacgccga gaacgccgtg atttttctgc atggtaacgc tgcctccagc
     1861 tacctgtgga ggcacgtcgt gcctcacatc gagcccgtgg ctagatgcat catccctgat
     1921 ctgatcggaa tgggtaagtc cggcaagagc gggaatggct catatcgcct cctggatcac
     1981 tacaagtacc tcaccgcttg gttcgagctg ctgaaccttc caaagaaaat catctttgtg
     2041 ggccacgact ggggggcttg tctggccttt cactactcct acgagcacca agacaagatc
     2101 aaggccatcg tccatgctga gagtgtcgtg gacgtgatcg agtcctggga cgagtggcct
     2161 gacatcgagg aggatatcgc cctgatcaag agcgaagagg gcgagaaaat ggtgcttgag
     2221 aataacttct tcgtcgagac catgctccca agcaagatca tgcggaaact ggagcctgag
     2281 gagttcgctg cctacctgga gccattcaag gagaagggcg aggttagacg gcctaccctc
     2341 tcctggcctc gcgagatccc tctcgttaag ggaggcaagc ccgacgtcgt ccagattgtc
     2401 cgcaactaca acgcctacct tcgggccagc gacgatctgc ctaagatgtt catcgagtcc
     2461 gaccctgggt tcttttccaa cgctattgtc gagggagcta agaagttccc taacaccgag
     2521 ttcgtgaagg tgaagggcct ccacttcagc caggaggacg ctccagatga aatgggtaag
     2581 tacatcaaga gcttcgtgga gcgcgtgctg aagaacgagc agtaattcta gataagtcca
     2641 actactaaac tggggattcc tgggccctga agaagggccc ctcgactaag tccaactact
     2701 aaactgggcc ctgaagaagg gcccatatag ggccctgaag aagggcccta tcgaggatat
     2761 tatctcgact aagtccaact actaaactgg gccctgaaga agggcccata tagggccctg
     2821 aagaagggcc ctatcgagga tattatctcg aggcggccgc ggccgcttcg agcagacatg
     2881 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2941 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3001 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3061 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc aggtggcact
     3121 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3181 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3241 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3301 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3361 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3421 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3481 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3541 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3601 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3661 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3721 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3781 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3841 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3901 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3961 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4021 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4081 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4141 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4201 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4261 aaaggatctt c
//
