LOCUS       sequence-343188-        4020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for making Nluc mRNA reporter with pA63..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4020)
  AUTHORS   Xiang K, Bartel DP
  TITLE     The molecular basis of coupling between poly(A)-tail length and 
            translational efficiency.
  JOURNAL   Elife. 2021 Jul 2;10. pii: 66493. doi: 10.7554/eLife.66493.
  PUBMED    34213414
REFERENCE   2  (bases 1 to 4020)
  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: "Elife."; 
            date: "2021-07-2"; volume: "10. pii: 66493. doi"; pages: " 
            10.7554/eLife.66493"
COMMENT     Sequence Label: C093
FEATURES             Location/Qualifiers
     source          1..4020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           16..58
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             70..585
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     3'UTR           585..710
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(891..909)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(892..909)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(927..943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(927..943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(940..962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    951..967
                     /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(975..981,982..999,1000..1005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1020..1041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1158..1175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1329..1917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1409..1428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2088..2879,2880..2948))
                     /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     2711..2730
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2949..3053)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3121..3139
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3177..3199)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3299..3318
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3385..3840)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3522..3543)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3734..3753
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3966..3988
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3980..3997
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3981..3997
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4004..4020,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4004..4020,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaataca agcttcttgt tctttttgca gaagctcaga ataaacgctc aactttggca
       61 gatctgaaca tggtcttcac actcgaagat ttcgttgggg actggcgaca gacagccggc
      121 tacaacctgg accaagtcct tgaacaggga ggtgtgtcca gtttgtttca gaatctcggg
      181 gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg aaaatgggct gaagatcgac
      241 atccatgtca tcatcccgta tgaaggtctg agcggcgacc aaatgggcca gatcgaaaaa
      301 atttttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct gcactatggc
      361 acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggacg gccgtatgaa
      421 ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac
      481 aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatc
      541 aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cgtaaaccag cctcaagaac
      601 acccgaatgg agtctctaag ctacataata ccaacttaca ctttacaaaa tgttgtcccc
      661 caaaatgtag ccattcgtat ctgctcctaa taaaaagaaa gtttcttcac attctggcgt
      721 agatccagac atgaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
      781 aaaaaaaaaa aaaaaagcta gccatggtcc cagcctcctc gctggcggct agtgggcaac
      841 atgcttcggc atggcgaatg ggacgcggcc gcatgcataa gcttgagtat tctatagtgt
      901 cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
      961 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1021 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1081 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1141 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1201 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1261 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1321 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1381 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1441 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1501 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1561 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1621 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1681 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1741 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1801 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1861 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1921 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1981 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2041 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2101 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2161 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2221 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2281 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2341 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2401 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2461 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2521 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2581 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2641 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2701 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2761 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2821 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2881 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2941 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3001 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3061 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3121 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3181 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     3241 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     3301 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     3361 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt
     3421 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3481 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3541 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3601 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3661 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3721 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3781 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3841 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3901 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3961 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata
//
