LOCUS       sequence-211069-        4536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monitor the expression of Luciferase driven by a mutated NRAS 
            5'-UTR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4536)
  AUTHORS   Kumari S, Bugaut A, Huppert JL, Balasubramanian S
  TITLE     An RNA G-quadruplex in the 5' UTR of the NRAS proto-oncogene 
            modulates translation.
  JOURNAL   Nat Chem Biol. 2007 Apr;3(4):218-21. doi: 10.1038/nchembio864. Epub 
            2007 Feb 25.
  PUBMED    17322877
REFERENCE   2  (bases 1 to 4536)
  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: 
            "10.1038/nchembio864"; journalName: "Nat Chem Biol"; date: 
            "2007-04"; volume: "3"; issue: "4"; pages: "218-21"
COMMENT     Sequence Label: pSKC12 - DelQ
FEATURES             Location/Qualifiers
     source          1..4536
                     /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"
     primer_bind     405..424
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        405..423
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             647..2299
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(713..731)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2315..2331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2315..2331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2328..2350)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2339..2355
                     /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(2363..2369,2370..2387,2388..2393))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2408..2429
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2546..2563)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2717..3305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2797..2816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3476..4267,4268..4336))
                     /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     4099..4118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4337..4441)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4509..4527
                     /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 ccagtgccaa gctttaatac gactcactat
      421 agggtgcggc ctgccgcatg actcgtggtt cggaggccca cgtggccggg gcggggactc
      481 aggcgcctgg ggcgccgact gattacgtag cgggcggggc cggaagtgcc gctccttggt
      541 gggggctgtt catggcggtt ccggggtctc caacattttt cccggctgtg gtcctaaatc
      601 tgtccaaagc agaggcagtg gagcttgagg ttcttgctgg tgtgaaatgg aagacgccaa
      661 aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca
      721 actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc
      781 acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga
      841 agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc
      901 tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc
      961 gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt
     1021 ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat
     1081 catccaaaaa attattatca tggattctaa aacggattac cagggatttc agtcgatgta
     1141 cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc
     1201 cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc
     1261 taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc
     1321 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     1381 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     1441 gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc
     1501 gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga
     1561 tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga
     1621 agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac
     1681 tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt
     1741 tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa
     1801 tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc
     1861 ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag acatagctta
     1921 ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa
     1981 aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt
     2041 cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt
     2101 tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca
     2161 agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg
     2221 tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg
     2281 cggaaagatc gccgtgtaag aattcgtaat catggtcata gctgtttcct gtgtgaaatt
     2341 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     2401 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2461 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2521 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2581 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2641 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2701 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2761 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2821 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2881 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2941 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3001 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3061 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3121 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3181 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3241 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3361 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3421 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3541 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3661 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3721 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3901 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4021 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4081 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4141 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4201 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4321 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4381 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4441 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4501 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
