LOCUS       sequence-340999-        7272 bp    DNA     circular SYN 14-APR-2025
DEFINITION  SCAT3.2 is fluorescent reporter (FRET probe) for cell 
            survival/apoptosis with Caspase-3 sensitive sequence 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 7272)
  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 7272)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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- SCAT3.2
FEATURES             Location/Qualifiers
     source          1..7272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..43
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     59..75
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    101..475
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     413..429
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      504..513
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             510..1196
                     /codon_start=1
                     /product="Anemonia majano cyan fluorescent protein"
                     /label=AmCyan1
                     /note="mammalian codon-optimized"
                     /translation="MALSNKFIGDDMKMTYHMDGCVNGHYFTVKGEGSGKPYEGTQTST
                     FKVTMANGGPLAFSFDILSTVFMYGNRCFTAYPTSMPDYFKQAFPDGMSYERTFTYEDG
                     GVATASWEISLKGNCFEHKSTFHGVNFPADGPVMAKKTTGWDPSFEKMTVCDGILKGDV
                     TAFLMLQGGGNYRCQFHTSYKTKKPVTMPPNHVVEHRIARTDLDKGGNSVQLTEHAVAH
                     ITSVVPF"
     CDS             join(1251..1253,1254..1967)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1293..1314)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1554..1573)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1901..1922
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(1998..2572,2573..2575,2576..2584)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2165..2182)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2392..2411
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2585..3187
                     /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     3084..3104
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3358..3827
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3957..3974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4128..4716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4208..4227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4880..5671,5672..5740))
                     /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     5503..5522
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5741..5845)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5875..6468
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6531..6888
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             join(6947..7272,1..91)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
       61 ttatccagcc ctcactcctt ctctaggcgc ccccatatgg ccatatgaga tcttatatgg
      121 ggcacccccg ccccttgtaa acttccctga ccctgacatg acaagagtta ctaacagccc
      181 ctctctccaa gctcacttac aggctctcta cttagtccag cacgaagtct ggagacctct
      241 ggcggcagcc taccaagaac aactggaccg accggtggta cctcaccctt accgagtcgg
      301 cgacacagtg tgggtccgcc gacaccagac taagaaccta gaacctcgct ggaaaggacc
      361 ttacacagtc ctgctgacca cccccaccgc cctcaaagta gacggcatcg cagcttggat
      421 acacgccgcc cacgtgaagg ctgccgaccc cgggggtgga ccatcctcta gactgccgga
      481 tctagctagt taattaagga tccgccacca tggccctgtc caacaagttc atcggcgacg
      541 acatgaagat gacctaccac atggacggct gcgtgaacgg ccactacttc accgtgaagg
      601 gcgagggcag cggcaagccc tacgagggca cccagacctc caccttcaag gtgaccatgg
      661 ccaacggcgg ccccctggcc ttctccttcg acatcctgtc caccgtgttc atgtacggca
      721 accgctgctt caccgcctac cccaccagca tgcccgacta cttcaagcag gccttccccg
      781 acggcatgtc ctacgagaga accttcacct acgaggacgg cggcgtggcc accgccagct
      841 gggagatcag cctgaagggc aactgcttcg agcacaagtc caccttccac ggcgtgaact
      901 tccccgccga cggccccgtg atggccaaga agaccaccgg ctgggacccc tccttcgaga
      961 agatgaccgt gtgcgacggc atcttgaagg gcgacgtgac cgccttcctg atgctgcagg
     1021 gcggcggcaa ctacagatgc cagttccaca cctcctacaa gaccaagaag cccgtgacca
     1081 tgccccccaa ccacgtggtg gagcaccgca tcgccagaac cgacctggac aagggcggca
     1141 acagcgtgca gctgaccgag cacgccgtgg cccacatcac ctccgtggtg cccttctcct
     1201 cgtccgagct cagcggagat gaggtcgatg gtaccagcgg aagcgaattc gtgagcaagg
     1261 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1321 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1381 tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1441 tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1501 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1561 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1621 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1681 actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     1741 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     1801 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     1861 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1921 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagcg gccgctacgt
     1981 aaattccgcc cccccccccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     2041 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     2101 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     2161 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     2221 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     2281 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     2341 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     2401 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     2461 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     2521 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     2581 aaccatggtt accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     2641 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     2701 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     2761 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     2821 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     2881 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     2941 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     3001 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     3061 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     3121 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     3181 tgcctgagtc gacgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     3241 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     3301 aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg
     3361 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     3421 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3481 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     3541 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     3601 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     3661 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac
     3721 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     3781 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta
     3841 tcaaaattaa tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa
     3901 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3961 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4021 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4081 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4141 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4201 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4261 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4321 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4381 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4441 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4501 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4561 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4621 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4681 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4741 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4801 aaaaggatct tcacctagat ccttttgcgg ccggccgcaa atcaatctaa agtatatatg
     4861 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4921 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4981 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5041 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5101 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5161 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5221 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5281 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5341 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5401 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5461 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5521 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5581 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5641 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5701 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5761 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5821 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gcataatgaa
     5881 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     5941 aaatacataa ctgagaatag aaaagttcag atcaaggtca ggaacagatg gaacagctga
     6001 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     6061 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     6121 ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa
     6181 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     6241 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     6301 gagcccacaa cccctcactc ggcgcgccag tcctccgatt gactgagtcg cccgggtacc
     6361 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     6421 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc
     6481 cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt aagctggcca
     6541 gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat gcgcctgcgt
     6601 cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc
     6661 ggaacacccg gccgcaaccc tgggagacgt cccagggact tcgggggccg tttttgtggc
     6721 ccgacctgag tccaaaaatc ccgatcgttt tggactcttt ggtgcacccc cctaatagga
     6781 gggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt
     6841 tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt gtctgctgca gcatcgttct
     6901 gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac
     6961 cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca
     7021 gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt
     7081 taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat
     7141 caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg
     7201 ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc
     7261 gcctcctctt cc
//
