LOCUS       sequence-98996-e        8004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse NrasG12D was amplified and mutated by PCR from IMAGE clone 
            6475312 (Invitrogen) and cloned downstream from the IRES; 
            subsequently Luciferase was cloned upstream of the IRES.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8004)
  AUTHORS   Zuber J, Radtke I, Pardee TS, Zhao Z, Rappaport AR, Luo W, McCurrach
            ME, Yang MM, Dolan ME, Kogan SC, Downing JR, Lowe SW
  TITLE     Mouse models of human AML accurately predict chemotherapy response.
  JOURNAL   Genes Dev. 2009 Apr 1;23(7):877-89. doi: 10.1101/gad.1771409.
  PUBMED    19339691
REFERENCE   2  (bases 1 to 8004)
  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: "10.1101/gad"; 
            journalName: "Genes Dev"; date: "2009-04-1- 1"; volume: "23"; issue:
            "7"; pages: "877-89"
COMMENT     Sequence Label: MSCV-Luciferase-IRES-NrasG12D
FEATURES             Location/Qualifiers
     source          1..8004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1419..3071
                     /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(1485..1503)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    3150..3717
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3167..3719
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3311..3328)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3538..3557
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     LTR             4366..4880
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(5049..5065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5049..5065)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5062..5084)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5073..5089
                     /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(5097..5103,5104..5121,5122..5127))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5142..5163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5280..5297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5451..6039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5531..5550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6210..7001,7002..7070))
                     /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     6833..6852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7071..7175)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7243..7261
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7299..7321)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7421..7440
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7634..7656
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7784..7803
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8003..8004,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatccaccat ggaagacgcc aaaaacataa
     1441 agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata
     1501 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
     1561 aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga
     1621 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
     1681 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
     1741 tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg
     1801 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa
     1861 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     1921 tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata
     1981 gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg
     2041 tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg
     2101 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
     2161 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     2221 ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg
     2281 tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     2341 atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg
     2401 ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag
     2461 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     2521 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag
     2581 gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     2641 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     2701 aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc
     2761 aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag
     2821 gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     2881 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     2941 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     3001 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga
     3061 tcgccgtgta attctaggat ccgtcgacct gcagccatgc attgattcaa aattttccga
     3121 tcacgagact agcctcgagg ttaacgaatt ccgccccccc cccctaacgt tactggccga
     3181 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     3241 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     3301 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     3361 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     3421 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     3481 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     3541 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     3601 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     3661 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     3721 ggccacaacc atgactgagt acaaactggt ggtggttgga gcagatggtg ttgggaaaag
     3781 cgccttgacg atccagctaa tccagaacca ctttgtggat gaatatgatc ccaccataga
     3841 ggattcttac cgaaagcaag tggtgattga tggtgagacc tgcctgctgg acatactgga
     3901 cacagctgga caagaggagt acagtgccat gagagaccag tacatgagga caggcgaagg
     3961 gttcctctgt gtatttgcca tcaataatag caaatcattt gcagatatta acctctacag
     4021 ggagcaaatt aagcgtgtga aagattctga tgatgtcccc atggtgctgg taggcaacaa
     4081 gtgtgacttg ccaacaagga cagttgacac aaagcaagcc cacgaactgg ccaagagtta
     4141 cggaattcca ttcattgaga cctcagccaa gacccgacag ggtgtggagg atgcctttta
     4201 cacactggta agggagatac gccagtaccg aatgaaaaag ctcaacagca gtgacgatgg
     4261 cactcaaggt tgtatggggc tgccctgtgt gctgatgtag tcgacctgca gccaagctta
     4321 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     4381 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac
     4441 tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa
     4501 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     4561 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     4621 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     4681 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     4741 cagtcctccg atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     4801 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     4861 cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg
     4921 gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact
     4981 ccaatactcc tgaaatagtt cattatggac agcgcagaag agctggggag aattaattcg
     5041 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5101 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5161 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5881 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7201 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7261 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     7321 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     7381 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     7441 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     7501 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     7561 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     7621 acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca
     7681 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     7741 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     7801 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     7861 ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc
     7921 aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa aagattttat
     7981 ttagtctcca gaaaaagggg ggaa
//
