LOCUS       sequence-377360-        3728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription template plasmid for a minimal 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 3728)
  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 3728)
  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-min5UTR-hRluc-A60
FEATURES             Location/Qualifiers
     source          1..3728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        408..426
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      430..439
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             436..1371
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     primer_bind     complement(1507..1523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1507..1523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1520..1542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1531..1547
                     /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(1555..1561,1562..1579,1580..1585))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1600..1621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1738..1755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1909..2497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1989..2008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2668..3459,3460..3528))
                     /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     3291..3310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3529..3633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3701..3719
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctctaa tacgactcac
      421 tataggctag ccaccatggc ttccaaggtg tacgaccccg agcaacgcaa acgcatgatc
      481 actgggcctc agtggtgggc tcgctgcaag caaatgaacg tgctggactc cttcatcaac
      541 tactatgatt ccgagaagca cgccgagaac gccgtgattt ttctgcatgg taacgctgcc
      601 tccagctacc tgtggaggca cgtcgtgcct cacatcgagc ccgtggctag atgcatcatc
      661 cctgatctga tcggaatggg taagtccggc aagagcggga atggctcata tcgcctcctg
      721 gatcactaca agtacctcac cgcttggttc gagctgctga accttccaaa gaaaatcatc
      781 tttgtgggcc acgactgggg ggcttgtctg gcctttcact actcctacga gcaccaagac
      841 aagatcaagg ccatcgtcca tgctgagagt gtcgtggacg tgatcgagtc ctgggacgag
      901 tggcctgaca tcgaggagga tatcgccctg atcaagagcg aagagggcga gaaaatggtg
      961 cttgagaata acttcttcgt cgagaccatg ctcccaagca agatcatgcg gaaactggag
     1021 cctgaggagt tcgctgccta cctggagcca ttcaaggaga agggcgaggt tagacggcct
     1081 accctctcct ggcctcgcga gatccctctc gttaagggag gcaagcccga cgtcgtccag
     1141 attgtccgca actacaacgc ctaccttcgg gccagcgacg atctgcctaa gatgttcatc
     1201 gagtccgacc ctgggttctt ttccaacgct attgtcgagg gagctaagaa gttccctaac
     1261 accgagttcg tgaaggtgaa gggcctccac ttcagccagg aggacgctcc agatgaaatg
     1321 ggtaagtaca tcaagagctt cgtggagcgc gtgctgaaga acgagcagta attctaggcg
     1381 atcgctcgag cccgggaatt cgtttaaacc tagagcggcc gctggccgcc tgcagnaaaa
     1441 aaaaaaaaaa aaatgcatct ggtttcctgg tctgtctcct ctcggtttaa gcttggcgta
     1501 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1561 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1621 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1681 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1741 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1801 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2041 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2101 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2341 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2401 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2461 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2521 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2581 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2641 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2701 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2761 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2821 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2881 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2941 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3001 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3061 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3121 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3181 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3241 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3301 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3361 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3421 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3481 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3541 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3601 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3661 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     3721 ctttcgtc
//
