LOCUS       sequence-190190-        8393 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Retroviral vector expressing MEK1-IRES-Bsd-p2A-RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8393)
  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 8393)
  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: pMXs2-MEK1
FEATURES             Location/Qualifiers
     source          1..8393
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    3..570
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    20..572
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(164..181)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     391..410
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             584..976
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1004..1060
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1061..1774
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     LTR             2029..2498
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2628..2645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2799..3387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2879..2898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3551..4342,4343..4411))
                     /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     4174..4193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4412..4516)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4547..5063
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    5127..5484
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5543..5959
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5889..5911
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5927..5943
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5969..6343
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     6281..6297
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(7572..7574,7575..7577,7578..8288)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7572..7574,7575..7577,7578..8288)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7617..7638)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7878..7897)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8225..8246
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8301..8366
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
       61 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
      121 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
      181 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
      241 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
      301 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
      361 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
      421 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
      481 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
      541 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggttc ctttgtctca
      601 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
      661 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
      721 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
      781 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
      841 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
      901 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
      961 ttatgtgtgg gagggcggag gtggtggagc tagcggcagc ggcgccacca acttcagcct
     1021 gctgaagcag gccggcgacg tggaggagaa ccccggcccc atggtgtcta agggcgaaga
     1081 gctgattaag gagaacatgc acatgaagct gtatatggag ggcaccgtga acaaccacca
     1141 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
     1201 caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat
     1261 gtacggcagc agaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc
     1321 cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac
     1381 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
     1441 ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggccaa
     1501 caccgagatg ctgtaccccg ctgacggcgg cctggaaggc agaagcgaca tggccctgaa
     1561 gctcgtgggc gggggccacc tgatctgcaa cttcaagacc acatacagat ccaagaaacc
     1621 cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac cacagactgg aaagaatcaa
     1681 ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga
     1741 cctccctagc aaactggggc acaaacttaa ttgaacgcgt taagtcgacg ataaaataaa
     1801 agattttatt ttgtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     1861 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     1921 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
     1981 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     2041 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     2101 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     2161 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     2221 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     2281 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     2341 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     2401 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     2461 ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta
     2521 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc
     2581 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3121 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3181 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3241 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3301 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3361 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3421 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3481 ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt
     3541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcataatga aagaccccac
     4561 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     4621 actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     4681 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     4741 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     4801 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     4861 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
     4921 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     4981 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     5041 acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag
     5101 ggaccaccga cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat
     5161 tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc
     5221 tgtatctggc ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg
     5281 agacgtccca gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga
     5341 tcgttttgga ctctttggtg caccccccta ataggaggga tatgtggttc tggtaggaga
     5401 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag
     5461 ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     5521 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     5581 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     5641 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     5701 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     5761 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     5821 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     5881 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     5941 cactccttct ctaggcgccc ccatatggcc atatgagatc ttatatgggg cacccccgcc
     6001 ccttgtaaac ttccctgacc ctgacatgac aagagttact aacagcccct ctctccaagc
     6061 tcacttacag gctctctact tagtccagca cgaagtctgg agacctctgg cggcagccta
     6121 ccaagaacaa ctggaccgac cggtggtacc tcacccttac cgagtcggcg acacagtgtg
     6181 ggtccgccga caccagacta agaacctaga acctcgctgg aaaggacctt acacagtcct
     6241 gctgaccacc cccaccgccc tcaaagtaga cggcatcgca gcttggatac acgccgccca
     6301 cgtgaaggct gccgaccccg ggggtggacc atcctctaga ctgccggatc tagctagtta
     6361 attaaggatc cgccaccatg cccaagaaga agccgacgcc catccagctg aacccggccc
     6421 ccgacggctc tgcagttaac gggaccagct ctgcggagac caacttggag gccttgcaga
     6481 agaagctgga ggagctagag cttgatgagc agcagcgaaa gcgccttgag gcctttctta
     6541 cccagaagca gaaggtggga gaactgaagg atgacgactt tgagaagatc agtgagctgg
     6601 gggctggcaa tggcggtgtg gtgttcaagg tctcccacaa gccttctggc ctggtcatgg
     6661 ccagaaagct aattcatctg gagatcaaac ccgcaatccg gaaccagatc ataagggagc
     6721 tgcaggttct gcatgagtgc aactctccgt acatcgtggg cttctatggt gcgttctaca
     6781 gcgatggcga gatcagtatc tgcatggagc acatggatgg aggttctctg gatcaagtcc
     6841 tgaagaaagc tggaagaatt cctgaacaaa ttttaggaaa agttagcatt gctgtaataa
     6901 aaggcctgac atatctgatg gagaagcaca agatcatgca cagagatgtc aagccctcca
     6961 acatcctagt caactcccgt ggggagatca agctctgtga ctttggggtc agcgggcagc
     7021 tcatcgactc catggccaac tccttcgtgg gcacaaggtc ctacatgtcg ccagaaagac
     7081 tccaggggac tcattactct gtgcagtcag acatctggag catgggactg tctctggtag
     7141 agatggcggt tgggaggtat cccatccctc ctccagatgc caaggagctg gagctgatgt
     7201 ttgggtgcca ggtggaagga gatgcggctg agaccccacc caggccaagg acccccggga
     7261 ggccccttag ctcatatgga atggacagcc gacctcccat ggcaattttt gagttgttgg
     7321 attacatagt caacgagcct cctccaaaac tgcccagtgg agtgttcagt ctggaatttc
     7381 aagattttgt gaataaatgc ttaataaaaa accccgcaga gagagcagat ttgaagcaac
     7441 tcatggttca tgcttttatc aagagatctg atgctgagga agtggatttt gcaggttggc
     7501 tctgctccac catcggcctt aaccagccca gcacaccaac ccatgctgct ggcgtctaag
     7561 cggccgcagg catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     7621 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     7681 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     7741 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     7801 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     7861 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     7921 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     7981 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     8041 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     8101 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     8161 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     8221 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     8281 tgtacaaggg cggaggtggc gactacaaag accatgacgg tgattataaa gatcatgaca
     8341 ttgattacaa ggatgacgat gacaagtaat aataaattcc gccccccccc ccc
//
