LOCUS       sequence-440839-        6300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NRAS G12V/D38A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6300)
  AUTHORS   Odawara T, Yamauchi S, Ichijo H
  TITLE     Apoptosis signal-regulating kinase 1 promotes inflammation in 
            senescence and aging.
  JOURNAL   Commun Biol. 2024 Jun 5;7(1):691. doi: 10.1038/s42003-024-06386-0.
  PUBMED    38839869
REFERENCE   2  (bases 1 to 6300)
  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: "Commun 
            Biol."; date: "2024-06-5"; volume: "7(1):691. doi"; pages: " 
            10.1038/s42003-024-06386-0"
COMMENT     Sequence Label: pT/Caggs-NRAS G12V/D38A
FEATURES             Location/Qualifiers
     source          1..6300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..658
                     /label=chicken beta-actin promoter
     intron          659..1675
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1599..1621
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1683..1702
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        complement(1748..1766)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1749..1766)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1800..1817
                     /codon_start=1
                     /product="epitope tag from middle T antigen of mouse 
                     polyoma virus"
                     /label=Glu-Glu tag
                     /translation="EYMPME"
     primer_bind     complement(2395..2412)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2401..2625
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3079..3099)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3079..3097)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3118..3134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3118..3134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3131..3153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3142..3158
                     /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(3166..3172,3173..3190,3191..3196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3211..3232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3349..3366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3520..4108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3600..3619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4279..5070,5071..5139))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4902..4921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5140..5244)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5271..5726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5358..5377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5568..5589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5852..5874
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5866..5883
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5867..5883
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5893..5912
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5893..5911
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(join(6285..6300,1))
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(6286..6300,1..2))
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      841 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
      901 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     1021 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     1081 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
     1321 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     1381 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     1441 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     1561 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1621 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     1681 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcc tcgagggggg
     1741 gccctattct atagtgtcac ctaaatgcta gagctcgctg atccaccatg gaggaagagg
     1801 aatatatgcc tatggaaatg actgagtaca aactggtggt ggttggagca gttggtgttg
     1861 ggaaaagcgc actgacaatc cagctaatcc agaaccactt tgtagatgaa tatgatccca
     1921 ccatagaggc ttcttacaga aaacaagtgg ttatagatgg tgaaacctgt ttgttggaca
     1981 tactggatac agctggacaa gaagagtaca gtgccatgag agaccaatac atgaggacag
     2041 gcgaaggctt cctctgtgta tttgccatca ataatagcaa gtcatttgcg gatattaacc
     2101 tctacaggga acagattaag cgagtaaaag actcggatga tgtacctatg gtgctagtgg
     2161 gaaacaagtg tgatttgcca acaaggacag ttgatacaaa acaagcccac gaactggcca
     2221 agagttacgg gattccattc attgaaacct cagccaagac cagacagggt gttgaagatg
     2281 ctttttacac actggtaaga gaaatacgcc agtaccgaat gaaaaaactc aacagcagtg
     2341 atgatgggac tcagggttgt atgggattgc catgtgtggt gatgtaagga tcagcctcga
     2401 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2461 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2521 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2581 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2641 gaaccagcca tcacaaagct ctgacctcaa tcctatagaa aggaggaatg agccaaaatt
     2701 cacccaactt attgtgggaa gcttgtggaa ggctactcga aatgtttgac ccaagttaaa
     2761 caatttaaag gcaatgctac caaatactaa ttgagtgtat gtaaacttct gacccactgg
     2821 gaatgtgatg aaagaaataa aagctgaaat gaatcattct ctctactatt attctgatat
     2881 ttcacattct taaaataaag tggtgatcct aactgaccta agacagggaa tttttactag
     2941 gattaaatgt caggaattgt gaaaaagtga gtttaaatgt atttggctaa ggtgtatgta
     3001 aacttccgac ttcaactgta tagggatcct ctagctagag tcgacctcga gggggggccc
     3061 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     3121 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3181 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3241 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3301 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3361 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3421 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3481 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3541 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3601 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3661 gcttaccgga tacctgtccg cctttctccc ttcgggaacc gtggcgcttt ctcatagctc
     3721 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3781 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3841 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3901 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3961 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4021 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4081 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4141 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4201 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4261 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4321 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4381 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4441 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4501 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4561 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4621 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4681 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaactt
     4741 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4801 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4861 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4921 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4981 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5041 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5101 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5161 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5221 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     5281 tagcggcgca ttaagcgcgg cgggagtggt ggttacgcgc agcgtgaccg ctacacttgc
     5341 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5401 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5461 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5521 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5581 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5641 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5701 taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa
     5761 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     5821 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     5881 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg agctcggatc cctatacagt
     5941 tgaagtcgga agtttacata cacttaagtt ggagtcatta aaactcgttt ttcaactact
     6001 ccacaaattt cttgttaaca aacaatagtt ttggcaagtc agttaggaca tctactttgt
     6061 gcatgacaca agtcattttt ccaacaattg tttacagaca gattatttca cttataattc
     6121 actgtatcac aattccagtg ggtcagaagt ttacatacac taagttgact gtgcctttaa
     6181 acagcttgga aaattccaga aaatgatgtc atggctttag aagctagatc tggccatcta
     6241 gagcggccgc gcgcactagt gaattccatg gatatcaagc ttatcgatac cgtcgacctc
//
