LOCUS       sequence-436826-        3259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NRAS-5'UTR-NanoLuc mRNA synthesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3259)
  TITLE     Protein purification and mRNA synthesis plasmids
REFERENCE   2  (bases 1 to 3259)
  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"
COMMENT     Sequence Label: pUC57_NRAS-5'UTR_Nanoluc
FEATURES             Location/Qualifiers
     source          1..3259
                     /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     402..421
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        402..420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      571..580
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1169..1185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1169..1185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1182..1204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1193..1209
                     /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(1217..1223,1224..1241,1242..1247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1262..1283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1400..1417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1571..2159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1651..1670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2337..3146)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3060..3079
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3138..3157
                     /label=pENTR-R
                     /note="pENTR vectors, reverse 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 ccagagaatt ctaatacgac tcactatagg
      421 gagagagctc ggggccggaa gtgccgctcc ttggtggggg ctgttcatgg cggttccggg
      481 gtctccaaca tttttcccgg ctgtggtcct aaatctgtcc aaagcagagg cagtggagct
      541 tgaggttctt gctggtgtga actgcaggcc gccaccatgg tttttacttt agaagatttt
      601 gttggtgatt ggcgtcaaac tgctggttat aatttagatc aagttttaga acaaggtggt
      661 gtttcttctt tgtttcaaaa cttaggtgtt tctgttactc ctattcaacg tattgtttta
      721 tctggtgaaa atggtttgaa gattgatatt catgttatta ttccatatga aggtttgtct
      781 ggtgatcaaa tgggtcaaat tgaaaagatt ttcaaagttg tttatcctgt ggacgatcat
      841 catttcaaag ttattttgca ttatggtact ttagttattg atggtgttac tcctaatatg
      901 attgattatt ttggtcgtcc ttatgaaggt attgctgttt tcgatggtaa aaagatcact
      961 gttactggta ctttatggaa tggtaacaag attattgatg aacgtttgat taatcctgat
     1021 ggttctttat tatttcgtgt tactattaat ggtgttactg gttggcgttt atgtgaacgt
     1081 attttagctt aagcggccgc accggtgcta gctcgagact agtcgacaaa aaaaaaaaaa
     1141 aaaaaaaaaa aagcttggtg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2281 gaagttttaa atcaagccca atctgaataa tgttacaacc aattaaccaa ttctgattag
     2341 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     2401 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     2461 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     2521 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     2581 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     2641 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     2701 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     2761 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     2821 tctaatacct ggaatgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca
     2881 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     2941 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     3001 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta
     3061 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
     3121 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     3181 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     3241 gacacgggcc agagctgca
//
