LOCUS       sequence-390164-        7152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Mouse DsRed-Monomer CASP9 C325A Mutant Fusion Protein in 
            Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7152)
  AUTHORS   Han J, Goldstein LA, Hou W, Watkins SC, Rabinowich H
  TITLE     Involvement of CASP9 (caspase 9) in IGF2R/CI-MPR endosomal 
            transport.
  JOURNAL   Autophagy. 2021 Jun;17(6):1393-1409. doi: 
            10.1080/15548627.2020.1761742. Epub 2020 May 25.
  PUBMED    32397873
REFERENCE   2  (bases 1 to 7152)
  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"; doi: 
            "10.1080/15548627.2020.1761742"; journalName: "Autophagy"; date: 
            "2021-06"; volume: "17"; issue: "6"; pages: "1393-1409"
COMMENT     Sequence Label: Mouse DsRed-Monomer CASP9 C325A Mutant
FEATURES             Location/Qualifiers
     source          1..7152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        147..450
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        451..654
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     605..625
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      693..702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             699..1373
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /label=DsRed-Monomer
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
                     VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
                     ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
                     GSQ"
     primer_bind     1097..1116
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2967..3088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3058..3077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3095..3550)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3232..3253)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3444..3463
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3577..3681
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3683..4040
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3683..3874
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3706..3726)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3891..4026
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4048..5073
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4048..5073
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5190..5271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5299..5317)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5385..5489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5490..5558,5559..6350)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5708..5727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6536..7124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7025..7044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
       61 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
      121 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      181 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      241 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      301 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      361 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      421 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      481 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      541 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      601 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      661 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggacaacacc gaggacgtca
      721 tcaaggagtt catgcagttc aaggtgcgca tggagggctc cgtgaacggc cactacttcg
      781 agatcgaggg cgagggcgag ggcaagccct acgagggcac ccagaccgcc aagctgcagg
      841 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg
      901 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacatgaag ctgtccttcc
      961 ccgagggctt cacctgggag cgctccatga acttcgagga cggcggcgtg gtggaggtgc
     1021 agcaggactc ctccctgcag gacggcacct tcatctacaa ggtgaagttc aagggcgtga
     1081 acttccccgc cgacggcccc gtaatgcaga agaagactgc cggctgggag ccctccaccg
     1141 agaagctgta cccccaggac ggcgtgctga agggcgagat ctcccacgcc ctgaagctga
     1201 aggacggcgg ccactacacc tgcgacttca agaccgtgta caaggccaag aagcccgtgc
     1261 agctgcccgg caaccactac gtggactcca agctggacat caccaaccac aacgaggact
     1321 acaccgtggt ggagcagtac gagcacgccg aggcccgcca ctccggctcc cagtccggac
     1381 tcagatctcg agctcaagct tcgaattctg cagtcgacat ggacgaggcg gaccggcagc
     1441 tcctgcggcg atgcagggtg cgcctagtga gcgagctgca agtcgcggag ctctgggacg
     1501 ctctgctgag tcgagagctc ttcacgcgcg acatgatcga ggatattcag caggcaggat
     1561 ctgggtctcg gcgggatcag gccaggcagc tggtcacaga ccttgagacc cgagggaggc
     1621 aggcccttcc tctcttcatc tcctgcttag aggacacagg ccaaggcacc ctggcttcac
     1681 tcttgcaaag cggtcggcaa gcagccaagc aggatccaga ggctgttaaa cccctagacc
     1741 acctggtgcc tgtggtcctg ggaccaatgg gactcacagc aaaggagcag agagtagtga
     1801 agctggaccc gtcacagcct gccgtgggaa acctcacccc agtggtgctg gggccagaag
     1861 agctctggcc tgctcggctc aagccagagg ttctcagacc agaaacaccc aggcccgtgg
     1921 acattggttc tggcggagct catgatgtct gtgttccagg gaagatcagg ggacatgcag
     1981 atatggcata caccctggat tcggatccct gtggccactg cctcatcatc aacaatgtga
     2041 acttctgccc ttcctcgggg ctcggcacac gcacgggctc caacttggac cgtgacaaac
     2101 ttgagcaccg attccgctgg ctgcgcttca tggtggaggt gaagaacgac ctgactgcca
     2161 agaaaatggt cacggctttg atggagatgg cacaccggaa ccaccgtgcc ctggactgct
     2221 ttgtggtggt catcctctct catggctgcc aggccagcca cctccagttc ccgggtgctg
     2281 tctatgggac agatggatgc tccgtgtcca ttgagaaaat tgtgaatatc ttcaacggga
     2341 gcggctgccc cagcctggga gggaagccca agctcttctt catccaggcc gcgggtggtg
     2401 agcagaaaga ccatggcttt gaggtggcct gcacttcctc tcaaggcagg acctcggaca
     2461 gtgactctga gccagatgct gtcccctatc aggaaggccc aaggcccttg gaccagctgg
     2521 atgctgtgtc aagtttgcct acccccagtg acatccttgt gtcctactcc accttcccag
     2581 gttttgtctc ctggagggac aagaaaagtg gctcctggta catcgagacc ttggatggca
     2641 ttctggagca gtgggctcgc tctgaagacc tgcagtccct ccttctcagg gttgccaatg
     2701 ctgtttctgc gaaagggact tacaagcaga ttcctggctg ttttaacttc ctccggaaaa
     2761 agctgttttt taaaacttca tgaggctcgg ggctgctctc tgcctcatta ttctctccga
     2821 agggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca
     2881 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2941 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3061 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     3121 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3181 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3241 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3301 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3361 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3421 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3481 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3541 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3601 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3661 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3721 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3961 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4021 gaggcctagg cttttgcaaa gatcgatatg aaaaagcctg aactcaccgc gacgtctgtc
     4081 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     4141 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     4201 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     4261 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
     4321 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     4381 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     4441 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     4501 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     4561 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     4621 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     4681 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     4741 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     4801 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     4861 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     4921 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     4981 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     5041 cgccccagca ctcgtccgag ggcaaaggaa tagagttcta gaggatcata atcagccata
     5101 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     5161 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     5221 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cctcgcgatc
     5281 cacaaccatg gcgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5341 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5401 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5461 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5521 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5581 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5641 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5701 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     5761 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5821 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5881 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5941 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6001 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6061 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6121 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6181 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6241 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6301 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ggcgcgccag
     6361 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     6421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6661 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6901 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7021 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7081 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7141 cctttttacg gt
//
