LOCUS       sequence-357349-        7863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector expressing RAC1(G12V)-IRES-Bsd-p2A-RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7863)
  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 7863)
  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-RAC1 G12V
FEATURES             Location/Qualifiers
     source          1..7863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..594
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    657..1014
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1073..1489
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1419..1441
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1457..1473
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1499..1873
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1811..1827
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1902..1911
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1911..1976
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(2571..2573,2574..2576,2577..3287)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2571..2573,2574..2576,2577..3287)
                     /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(2616..2637)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2877..2896)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3224..3245
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3300..3365
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(3390..3964,3965..3967,3968..3976)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3557..3574)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3784..3803
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3977..4369
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4397..4453
                     /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             4454..5167
                     /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             5347..5816
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5946..5963)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6117..6705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6197..6216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6869..7660,7661..7729))
                     /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     7492..7511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7730..7834)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
      121 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      181 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      241 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
      301 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      361 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      421 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
      481 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
      541 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
      601 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      661 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
      721 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      781 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
      841 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta
      901 ataggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      961 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1021 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     1081 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1141 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1201 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1261 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1321 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1381 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1441 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
     1501 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     1561 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     1621 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     1681 tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga
     1741 acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga
     1801 cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc
     1861 atcctctaga ctgccggatc tagctagtta attaaggatc cgccaccatg gactataagg
     1921 accatgatgg agattataag gaccacgata ttgactataa agacgatgac gacaaagggg
     1981 gccaggcaat caagtgtgtg gtggttggag atgtagccgt gggcaagacc tgtttgctga
     2041 tttcttacac aaccaatgcg ttccccggcg aatacattcc gacagttttc gataactaca
     2101 gcgcaaatgt tatggtggac gggaaaccgg tgaatctggg actctgggat actgccggcc
     2161 aggaggacta cgatcgcctc cggccgctct cctatcctca gacagatgtg ttcctcatct
     2221 gcttctcact cgtatcccca gcaagtttcg agaacgttcg ggctaagtgg taccccgaag
     2281 tacgccacca ctgtcctaat accccaatca ttctcgtcgg gactaagctc gatctgcggg
     2341 atgataaaga cacgatcgag aaattgaagg agaaaaaact cacacctatc acataccccc
     2401 agggtctggc aatggctaag gagataggcg ctgtgaaata cttggagtgc tctgccctta
     2461 cacagagagg cctgaaaact gtgtttgatg aggccattcg agcagtcctg tgtccgcccc
     2521 ccgttaagaa gaggaaacgg aagtgtttgt tgctgtaggc ggccgcaggc atggtgagca
     2581 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2641 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2701 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2761 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2821 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2881 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2941 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3001 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     3061 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     3121 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     3181 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3241 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggaggtggcg
     3301 actacaaaga ccatgacggt gattataaag atcatgacat tgattacaag gatgacgatg
     3361 acaagtaata ataaattccg cccccccccc ccctctccct cccccccccc taacgttact
     3421 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     3481 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     3541 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     3601 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     3661 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     3721 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     3781 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     3841 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     3901 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     3961 taatatggcc acaaccatgg ttcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     4021 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     4081 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     4141 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     4201 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     4261 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     4321 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggcg gaggtggtgg
     4381 agctagcggc agcggcgcca ccaacttcag cctgctgaag caggccggcg acgtggagga
     4441 gaaccccggc cccatggtgt ctaagggcga agagctgatt aaggagaaca tgcacatgaa
     4501 gctgtatatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg
     4561 caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc
     4621 cttcgccttc gacatcctgg ctaccagctt catgtacggc agcagaacct tcatcaacca
     4681 cacccagggc atccccgact tctttaagca gtccttccct gagggcttca catgggagag
     4741 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
     4801 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt
     4861 gatgcagaag aaaacactcg gctgggaggc caacaccgag atgctgtacc ccgctgacgg
     4921 cggcctggaa ggcagaagcg acatggccct gaagctcgtg ggcgggggcc acctgatctg
     4981 caacttcaag accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt
     5041 ctactatgtg gaccacagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga
     5101 gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacaaact
     5161 taattgaacg cgttaagtcg acgataaaat aaaagatttt attttgtctc cagaaaaagg
     5221 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     5281 aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg
     5341 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     5401 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     5461 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
     5521 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     5581 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     5641 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
     5701 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     5761 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcacatg
     5821 cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa atggccatag
     5881 ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5941 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6001 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6061 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6121 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6181 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6241 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6301 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6361 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6421 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6481 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6541 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6601 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6661 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6721 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6781 agattatcaa aaaggatctt cacctagatc cttttgcggc cggccgcaaa tcaatctaaa
     6841 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6901 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6961 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7021 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7081 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7141 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7201 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7321 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7381 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7441 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7501 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7561 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7621 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7681 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7741 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7801 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7861 cat
//
