LOCUS       sequence-190191-        8409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector expressing MEK1-IRES-Bsd-p2A-RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8409)
  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 8409)
  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..8409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             23..539
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    603..960
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1019..1435
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1365..1387
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1403..1419
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1445..1819
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1757..1773
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(3048..3050,3051..3053,3054..3764)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3048..3050,3051..3053,3054..3764)
                     /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(3093..3114)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3354..3373)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3701..3722
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3777..3842
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(3882..4456,4457..4459,4460..4468)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4049..4066)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4276..4295
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4469..4861
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4889..4945
                     /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             4946..5659
                     /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             5914..6383
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6513..6530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6684..7272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6764..6783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7436..8227,8228..8296))
                     /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     8059..8078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8297..8401)
                     /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 tggtccccag atgcggtcca gccctcagca
      241 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
      301 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
      361 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga ttgactgagt
      421 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
      481 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
      541 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
      601 gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt
      661 atgcgcctgc gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
      721 actgacgagt tcggaacacc cggccgcaac cctgggagac gtcccaggga cttcgggggc
      781 cgtttttgtg gcccgacctg agtccaaaaa tcccgatcgt tttggactct ttggtgcacc
      841 cccctaatag gagggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
      901 tccgtctgaa tttttgcttt cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg
      961 cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg
     1021 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     1081 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     1141 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     1201 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     1261 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     1321 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     1381 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat
     1441 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga
     1501 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt
     1561 ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt
     1621 ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa
     1681 cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa
     1741 agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg
     1801 tggaccatcc tctagactgc cggatctagc tagttaatta aggatccgcc accatgccca
     1861 agaagaagcc gacgcccatc cagctgaacc cggcccccga cggctctgca gttaacggga
     1921 ccagctctgc ggagaccaac ttggaggcct tgcagaagaa gctggaggag ctagagcttg
     1981 atgagcagca gcgaaagcgc cttgaggcct ttcttaccca gaagcagaag gtgggagaac
     2041 tgaaggatga cgactttgag aagatcagtg agctgggggc tggcaatggc ggtgtggtgt
     2101 tcaaggtctc ccacaagcct tctggcctgg tcatggccag aaagctaatt catctggaga
     2161 tcaaacccgc aatccggaac cagatcataa gggagctgca ggttctgcat gagtgcaact
     2221 ctccgtacat cgtgggcttc tatggtgcgt tctacagcga tggcgagatc agtatctgca
     2281 tggagcacat ggatggaggt tctctggatc aagtcctgaa gaaagctgga agaattcctg
     2341 aacaaatttt aggaaaagtt agcattgctg taataaaagg cctgacatat ctgatggaga
     2401 agcacaagat catgcacaga gatgtcaagc cctccaacat cctagtcaac tcccgtgggg
     2461 agatcaagct ctgtgacttt ggggtcagcg ggcagctcat cgactccatg gccaactcct
     2521 tcgtgggcac aaggtcctac atgtcgccag aaagactcca ggggactcat tactctgtgc
     2581 agtcagacat ctggagcatg ggactgtctc tggtagagat ggcggttggg aggtatccca
     2641 tccctcctcc agatgccaag gagctggagc tgatgtttgg gtgccaggtg gaaggagatg
     2701 cggctgagac cccacccagg ccaaggaccc ccgggaggcc ccttagctca tatggaatgg
     2761 acagccgacc tcccatggca atttttgagt tgttggatta catagtcaac gagcctcctc
     2821 caaaactgcc cagtggagtg ttcagtctgg aatttcaaga ttttgtgaat aaatgcttaa
     2881 taaaaaaccc cgcagagaga gcagatttga agcaactcat ggttcatgct tttatcaaga
     2941 gatctgatgc tgaggaagtg gattttgcag gttggctctg ctccaccatc ggccttaacc
     3001 agcccagcac accaacccat gctgctggcg tctaagcggc cgcaggcatg gtgagcaagg
     3061 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     3121 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     3181 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3241 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     3301 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     3361 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     3421 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     3481 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     3541 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     3601 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     3661 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3721 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcgga ggtggcgact
     3781 acaaagacca tgacggtgat tataaagatc atgacattga ttacaaggat gacgatgaca
     3841 agtaataata aattccgccc cccccccccc cccccccccc ccccctctcc ctcccccccc
     3901 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     3961 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     4021 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     4081 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
     4141 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     4201 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     4261 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     4321 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
     4381 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     4441 aaaacacgat gataatatgg ccacaaccat ggttcctttg tctcaagaag aatccaccct
     4501 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     4561 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     4621 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     4681 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     4741 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     4801 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     4861 cggaggtggt ggagctagcg gcagcggcgc caccaacttc agcctgctga agcaggccgg
     4921 cgacgtggag gagaaccccg gccccatggt gtctaagggc gaagagctga ttaaggagaa
     4981 catgcacatg aagctgtata tggagggcac cgtgaacaac caccacttca agtgcacatc
     5041 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg
     5101 cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcagaac
     5161 cttcatcaac cacacccagg gcatccccga cttctttaag cagtccttcc ctgagggctt
     5221 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     5281 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc
     5341 caacggccct gtgatgcaga agaaaacact cggctgggag gccaacaccg agatgctgta
     5401 ccccgctgac ggcggcctgg aaggcagaag cgacatggcc ctgaagctcg tgggcggggg
     5461 ccacctgatc tgcaacttca agaccacata cagatccaag aaacccgcta agaacctcaa
     5521 gatgcccggc gtctactatg tggaccacag actggaaaga atcaaggagg ccgacaaaga
     5581 gacctacgtc gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact
     5641 ggggcacaaa cttaattgaa cgcgttaagt cgacgataaa ataaaagatt ttattttgtc
     5701 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     5761 cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt
     5821 caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     5881 gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg
     5941 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     6001 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     6061 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     6121 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     6181 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     6241 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     6301 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     6361 tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc
     6421 ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga
     6481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7381 ttgcggccgg ccgcaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7441 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7501 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7561 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7621 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7681 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7741 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7801 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7861 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7921 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7981 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8041 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8101 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8161 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8221 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8281 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8401 gcacatttc
//
