LOCUS       sequence-169427-        6560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector expressing NRAS(G13D)-IRES-RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6560)
  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 6560)
  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..6560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2521..2586
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(2614..3188,3189..3191,3192..3200)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2781..2798)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3008..3027
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3201..3896
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     LTR             4065..4534
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4664..4681)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4835..5423)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4915..4934)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5587..6378,6379..6447))
                     /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     6210..6229
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6448..6552)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggcc
     1921 accatgactg agtacaaact ggtggtggtt ggagcaggtg atgttgggaa aagcgcactg
     1981 acaatccagc taatccagaa ccactttgta gatgaatatg atcccaccat agaggattct
     2041 tacagaaaac aagtggttat agatggtgaa acctgtttgt tggacatact ggatacagct
     2101 ggacaagaag agtacagtgc catgagagac caatacatga ggacaggcga aggcttcctc
     2161 tgtgtatttg ccatcaataa tagcaagtca tttgcggata ttaacctcta cagggagcag
     2221 attaagcgag taaaagactc ggatgatgta cctatggtgc tagtgggaaa caagtgtgat
     2281 ttgccaacaa ggacagttga tacaaaacaa gcccacgaac tggccaagag ttacgggatt
     2341 ccattcattg aaacctcagc caagaccaga cagggtgttg aagatgcttt ttacacactg
     2401 gtaagagaaa tacgccagta ccgaatgaaa aaactcaaca gcagtgatga tgggactcag
     2461 ggttgtatgg gattgccatg tgtggtgatg taagcaggag gcggtggagg cggtgcaggc
     2521 gactacaaag accatgacgg tgattataaa gatcatgaca ttgattacaa ggatgacgat
     2581 gacaagtaat aataaattcc gccccccccc ccccccctct ccctcccccc cccctaacgt
     2641 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     2701 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     2761 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     2821 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     2881 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     2941 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     3001 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     3061 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     3121 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     3181 atgataatat ggccacaacc atgagcgagc tgatcaagga gaacatgcac atgaagctgt
     3241 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
     3301 cctacgaggg cacccagacc atgaagatca aggtggtcga gggcggccct ctccccttcg
     3361 ccttcgacat cctggctacc agcttcatgt acggcagcaa agccttcatc aaccacaccc
     3421 agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagaatca
     3481 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcttc cagaacggct
     3541 gcatcatcta caacgtcaag atcaacgggg tgaacttccc atccaacggc cctgtgatgc
     3601 agaagaaaac acgcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc
     3661 tgagaggcca cagccagatg gccctgaagc tcgtgggcgg gggctacctg cactgctcct
     3721 tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcttccact
     3781 tcgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
     3841 acgagatggc tgtggccaag tactgcgacc tccctagcaa actggggcac agataagtac
     3901 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     3961 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     4021 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     4081 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     4141 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4201 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     4261 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     4321 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     4381 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     4441 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     4501 gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt
     4561 ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac
     4621 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4681 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4741 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4801 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4861 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4921 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4981 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5041 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5101 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5161 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5221 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5281 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5341 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5401 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5461 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5521 cctagatcct tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc
     5581 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5641 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5701 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5761 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5821 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5881 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5941 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6001 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6061 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6121 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6181 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6241 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6301 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6361 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6421 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6481 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6541 aggggttccg cgcacatttc
//
