LOCUS       sequence-346537-        6825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mVenus-TOSI (TOr-Signal-Indicator) is a fluorescent reporter for 
            mTORC1 signaling (monitoring PDCD4 degradation) which can be 
            introduced to mammalian cells by retrovirus vector. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6825)
  AUTHORS   Oki T, Mercier F, Kato H, Jung Y, McDonald TO, Spencer JA, Mazzola 
            MC, van Gastel N, Lin CP, Michor F, Kitamura T, Scadden DT
  TITLE     Imaging dynamic mTORC1 pathway activity in vivo reveals marked 
            shifts that support time-specific inhibitor therapy in AML.
  JOURNAL   Nat Commun. 2021 Jan 11;12(1):245. doi: 10.1038/s41467-020-20491-8.
  PUBMED    33431855
REFERENCE   2  (bases 1 to 6825)
  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: "Nat 
            Commun."; date: "2021-01-11"; volume: "12(1):245. doi"; pages: " 
            10.1038/s41467-020-20491-8"
COMMENT     Sequence Label: pMXs-IP-mVenus-TOSI
FEATURES             Location/Qualifiers
     source          1..6825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..54
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(129..131,132..134,135..845)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(174..195)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(435..454)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     782..803
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             876..896
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    join(1176..1750,1751..1753,1754..1762)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1343..1360)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1570..1589
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1763..2365
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MVTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     2262..2282
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2536..3005
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3135..3152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3306..3894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3386..3405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4058..4849,4850..4918))
                     /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     4681..4700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4919..5023)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5053..5646
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5709..6066
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6125..6541
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6471..6493
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6509..6525
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6551..6825,1..100)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
ORIGIN
        1 gaccaccccc accgccctca aagtagacgg catcgcagct tggatacacg ccgcccacgt
       61 gaaggctgcc gaccccgggg gtggaccatc ctctagactg ccggatctag ctagttaatt
      121 aaggatccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      181 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      241 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
      301 ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc
      361 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      421 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      481 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      541 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
      601 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
      661 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      721 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      781 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      841 acaagggata tccatcacac tggcggccgg aattgccaaa aaagaagaga aaggtagaat
      901 tcatggatat agaaaatgag cagacactga atgtgaaccc cactgaccct gacaatttaa
      961 gcgactctct cttttctgga gatgaagaaa atgctggcac tgaagaaata aagaatgaaa
     1021 taaatggaaa ttggatttcc gcatctacta ttaatgaagc tagaatcaat gcaaaagcca
     1081 aaagacgact gcggaaaaat tcatcccggg actctggccg aggagactca gtcagtgaca
     1141 attgagcggc cgctacgtaa attccgcccc ccccccccct ctccctcccc cccccctaac
     1201 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     1261 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     1321 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     1381 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     1441 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     1501 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     1561 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     1621 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     1681 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     1741 cgatgataat atggccacaa ccatggttac cgagtacaag cccacggtgc gcctcgccac
     1801 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     1861 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     1921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     1981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     2041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     2101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     2161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     2221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     2281 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     2341 catgacccgc aagcccggtg cctgagtcga cgataaaata aaagatttta tttagtctcc
     2401 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     2461 cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag
     2521 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     2581 ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     2641 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc
     2701 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     2761 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     2821 ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg
     2881 actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt
     2941 ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc
     3001 tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa
     3061 tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3121 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3181 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3241 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3301 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3361 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3421 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3481 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3541 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3601 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3661 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3721 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3781 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3841 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3901 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3961 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc ggccgcaaat
     4021 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4081 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4141 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4201 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4261 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4321 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4381 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4441 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4501 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4561 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4621 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4681 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4741 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4801 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4861 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4921 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4981 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5041 tgccacctgc ataatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     5101 attttgcaag gcatggaaaa atacataact gagaatagaa aagttcagat caaggtcagg
     5161 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5221 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     5281 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     5341 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     5401 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     5461 tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc ctccgattga
     5521 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     5581 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     5641 ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac
     5701 cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgact
     5761 gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg gcggacccgt
     5821 ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc cagggacttc
     5881 gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg gactctttgg
     5941 tgcacccccc taataggagg gatatgtggt tctggtagga gacgagaacc taaaacagtt
     6001 cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg cgcgtcttgt
     6061 ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa
     6121 ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga
     6181 gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc
     6241 tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct
     6301 catcacccag gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt
     6361 cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt
     6421 tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc
     6481 tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc
     6541 ccccatatgg ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga
     6601 ccctgacatg acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta
     6661 cttagtccag cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg
     6721 accggtggta cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac
     6781 taagaaccta gaacctcgct ggaaaggacc ttacacagtc ctgct
//
