LOCUS       sequence-172761-        6529 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Retroviral vector expressing NRAS(G13D)-IRES-RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6529)
  AUTHORS   Wang T, Yu H, Hughes NW, Liu B, Kendirli A, Klein K, Chen WW, Lander
            ES, Sabatini DM
  TITLE     Gene Essentiality Profiling Reveals Gene Networks and Synthetic 
            Lethal Interactions with Oncogenic Ras.
  JOURNAL   Cell. 2017 Feb 1. pii: S0092-8674(17)30061-2. doi: 
            10.1016/j.cell.2017.01.013.
  PUBMED    28162770
REFERENCE   2  (bases 1 to 6529)
  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. 2017 
            Feb 1. pii: S0092-8674(17)30061-2. doi: 10.1016/j.cell.2017.01.013."
COMMENT     Sequence Label: pMXs3-NRAS G13D
FEATURES             Location/Qualifiers
     source          1..6529
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    13..565
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(157..174)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     384..403
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             577..1272
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     LTR             1441..1910
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2040..2057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2211..2799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2291..2310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2963..3754,3755..3823))
                     /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     3586..3605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3824..3928)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3958..4551
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4614..4971
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5030..5446
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5376..5398
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5414..5430
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5456..5830
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     5768..5784
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             6457..6522
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
       61 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
      121 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
      181 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
      241 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
      301 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
      361 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
      421 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
      481 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
      541 tcctttgaaa aacacgatga taatatggcc acaaccatga gcgagctgat caaggagaac
      601 atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc
      661 gagggcgaag gcaagcccta cgagggcacc cagaccatga agatcaaggt ggtcgagggc
      721 ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcaaagcc
      781 ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc tgagggcttc
      841 acatgggaga gaatcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc
      901 agcttccaga acggctgcat catctacaac gtcaagatca acggggtgaa cttcccatcc
      961 aacggccctg tgatgcagaa gaaaacacgc ggctgggagg ccaacaccga gatgctgtac
     1021 cccgctgacg gcggcctgag aggccacagc cagatggccc tgaagctcgt gggcgggggc
     1081 tacctgcact gctccttcaa gaccacatac agatccaaga aacccgctaa gaacctcaag
     1141 atgcccggct tccacttcgt ggaccacaga ctggaaagaa tcaaggaggc cgacaaagag
     1201 acctacgtcg agcagcacga gatggctgtg gccaagtact gcgacctccc tagcaaactg
     1261 gggcacagat aagtacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     1321 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     1381 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     1441 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     1501 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     1561 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     1621 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     1681 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     1741 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     1801 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     1861 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     1921 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     1981 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2041 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2101 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2161 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2221 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2281 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2341 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2401 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2461 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2521 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2581 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2641 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2701 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2761 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2821 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2881 tcaaaaagga tcttcaccta gatccttttg cggccggccg caaatcaatc taaagtatat
     2941 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3001 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3061 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3121 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3181 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3241 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3301 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3361 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3421 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3481 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3541 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3601 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3661 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3721 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3781 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3841 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3901 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgcataat
     3961 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     4021 gaaaaataca taactgagaa tagaaaagtt cagatcaagg tcaggaacag atggaacagc
     4081 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4141 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4201 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     4261 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     4321 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     4381 aaagagccca caacccctca ctcggcgcgc cagtcctccg attgactgag tcgcccgggt
     4441 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     4501 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc
     4561 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
     4621 ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgactgattt tatgcgcctg
     4681 cgtcggtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
     4741 ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg acttcggggg ccgtttttgt
     4801 ggcccgacct gagtccaaaa atcccgatcg ttttggactc tttggtgcac ccccctaata
     4861 ggagggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga
     4921 atttttgctt tcggtttggg accgaagccg cgccgcgcgt cttgtctgct gcagcatcgt
     4981 tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt
     5041 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     5101 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     5161 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     5221 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     5281 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
     5341 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
     5401 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccccca tatggccata
     5461 tgagatctta tatggggcac ccccgcccct tgtaaacttc cctgaccctg acatgacaag
     5521 agttactaac agcccctctc tccaagctca cttacaggct ctctacttag tccagcacga
     5581 agtctggaga cctctggcgg cagcctacca agaacaactg gaccgaccgg tggtacctca
     5641 cccttaccga gtcggcgaca cagtgtgggt ccgccgacac cagactaaga acctagaacc
     5701 tcgctggaaa ggaccttaca cagtcctgct gaccaccccc accgccctca aagtagacgg
     5761 catcgcagct tggatacacg ccgcccacgt gaaggctgcc gaccccgggg gtggaccatc
     5821 ctctagactg ccggatctag ctagttaatt aaggccacca tgactgagta caaactggtg
     5881 gtggttggag caggtgatgt tgggaaaagc gcactgacaa tccagctaat ccagaaccac
     5941 tttgtagatg aatatgatcc caccatagag gattcttaca gaaaacaagt ggttatagat
     6001 ggtgaaacct gtttgttgga catactggat acagctggac aagaagagta cagtgccatg
     6061 agagaccaat acatgaggac aggcgaaggc ttcctctgtg tatttgccat caataatagc
     6121 aagtcatttg cggatattaa cctctacagg gagcagatta agcgagtaaa agactcggat
     6181 gatgtaccta tggtgctagt gggaaacaag tgtgatttgc caacaaggac agttgataca
     6241 aaacaagccc acgaactggc caagagttac gggattccat tcattgaaac ctcagccaag
     6301 accagacagg gtgttgaaga tgctttttac acactggtaa gagaaatacg ccagtaccga
     6361 atgaaaaaac tcaacagcag tgatgatggg actcagggtt gtatgggatt gccatgtgtg
     6421 gtgatgtaag caggaggcgg tggaggcggt gcaggcgact acaaagacca tgacggtgat
     6481 tataaagatc atgacattga ttacaaggat gacgatgaca agtaataat
//
