LOCUS       sequence-465701-        7000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mutant form of NRAS .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7000)
  AUTHORS   Chen Z, Zeng C, Yang L, Che Y, Chen M, Sau L, Wang B, Zhou K, Chen 
            Y, Qing Y, Shen C, Zhang T, Wunderlich M, Wu D, Li W, Wang K, Leung 
            K, Sun M, Tang T, He X, Zhang L, Swaminathan S, Mulloy JC, Muschen 
            M, Huang H, Weng H, Xiao G, Deng X, Chen J
  TITLE     YTHDF2 promotes ATP synthesis and immune evasion in B cell 
            malignancies.
  JOURNAL   Cell. 2024 Dec 12:S0092-8674(24)01324-2. doi: 
            10.1016/j.cell.2024.11.007.
  PUBMED    39694037
REFERENCE   2  (bases 1 to 7000)
  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: "Cell. 2024 
            Dec 12:S0092-8674(24)01324-2. doi: 10.1016/j.cell.2024.11.007."
COMMENT     Sequence Label: MSCV-NRAS(G12D)-Neo
FEATURES             Location/Qualifiers
     source          1..7000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     70..89
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     283..305
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     433..452
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             652..1168
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1232..1573
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1640..2056
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1986..2008
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2024..2040
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        2655..3154
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2690..2713)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3087..3106
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3168..3971
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3231..3250)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3841..3860
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4046..4560
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4722..4738
                     /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(4746..4752,4753..4770,4771..4776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4791..4812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4929..4946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5100..5688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5180..5199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5859..6650,6651..6719))
                     /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     6482..6501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6720..6824)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6892..6910
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6948..6970)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
       61 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
      121 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
      181 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      241 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      301 aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
      361 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
      421 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
      481 gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta aagacaggat
      541 atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta
      601 cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
      661 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat
      721 acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg
      781 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
      841 gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt
      901 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
      961 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     1021 ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
     1081 ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc tcagattgat
     1141 tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg agacccctgc
     1201 ccagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt tcgtgtctgt
     1261 ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc tgtactagtt
     1321 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct gaacacccgg
     1381 ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc cgacctgagg
     1441 aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag gagacgagaa
     1501 cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc gaagccgcgc
     1561 gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga ctgtgtttct
     1621 gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga ccttaggtca
     1681 ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga agagacgttg
     1741 ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc gagacggcac
     1801 ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg gcccgcatgg
     1861 acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg acccccctcc
     1921 ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat ccgccccgtc
     1981 tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc cagccctcac
     2041 tccttctcta ggcgccggaa ttcatgactg agtacaaact ggtggtggtt ggagcagatg
     2101 gtgttgggaa aagcgcactg acaatccagc taatccagaa ccactttgta gatgaatatg
     2161 atcccaccat agaggattct tacagaaaac aagtggttat agatggtgaa acctgtttgt
     2221 tggacatact ggatacagct ggacaagaag agtacagtgc catgagagac caatacatga
     2281 ggacaggcga aggcttcctc tgtgtatttg ccatcaataa tagcaagtca tttgcggata
     2341 ttaacctcta cagggagcag attaagcgag taaaagactc ggatgatgta cctatggtgc
     2401 tagtgggaaa caagtgtgat ttgccaacaa ggacagttga tacaaaacaa gcccacgaac
     2461 tggccaagag ttacgggatt ccattcattg aaacctcagc caagaccaga cagggtgttg
     2521 aagatgcttt ttacacactg gtaagagaaa tacgccagta ccgaatgaaa aaactcaaca
     2581 gcagtgatga tgggactcag ggttgtatgg gattgccatg tgtggtgatg taactcgaga
     2641 gatctaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     2701 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     2761 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     2821 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     2881 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     2941 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     3001 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
     3061 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     3121 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agccaatatg ggatcggcca
     3181 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3241 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3301 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3361 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3421 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3481 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3541 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3601 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3661 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3721 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3781 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3841 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3901 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3961 agttcttctg aggggatccg tcgacctgca gccaagctta tcgataaaat aaaagatttt
     4021 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     4081 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     4141 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     4201 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     4261 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     4321 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     4381 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
     4441 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     4501 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     4561 gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt
     4621 aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt
     4681 cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt
     4741 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4801 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4861 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4921 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4981 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5041 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5101 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5161 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5221 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5281 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5341 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5401 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5461 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5521 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5581 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5641 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5701 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5761 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5821 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5881 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5941 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6001 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6061 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6121 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6181 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6241 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6301 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6361 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6421 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6481 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6541 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6601 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6661 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6721 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6781 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6841 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6901 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6961 cagctcccgg agacggtcac agcttgtctg taagcggatg
//
