LOCUS       sequence-390261-        7016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human DsRed-Monomer CASP9 C287A Mutant Fusion Protein in 
            Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7016)
  TITLE     Human DsRed-Monomer CASP9 C287A Mutant
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7016)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Human DsRed-Monomer CASP9 C287A Mutant
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        392..595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     546..566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      634..643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             640..1314
                     /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     1038..1057
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(2089..2100)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    2772..2893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2863..2882
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2900..3355)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3037..3058)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3249..3268
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3382..3486
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3488..3845
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3488..3679
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3511..3531)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3696..3831
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3853..4878
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3853..4878
                     /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    4995..5076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5104..5122)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5190..5294
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5295..5363,5364..6155)
                     /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(5513..5532)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6341..6929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6830..6849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      361 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      421 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      481 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      541 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      601 agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggacaacac cgaggacgtc
      661 atcaaggagt tcatgcagtt caaggtgcgc atggagggct ccgtgaacgg ccactacttc
      721 gagatcgagg gcgagggcga gggcaagccc tacgagggca cccagaccgc caagctgcag
      781 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttccagtac
      841 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacatgaa gctgtccttc
      901 cccgagggct tcacctggga gcgctccatg aacttcgagg acggcggcgt ggtggaggtg
      961 cagcaggact cctccctgca ggacggcacc ttcatctaca aggtgaagtt caagggcgtg
     1021 aacttccccg ccgacggccc cgtaatgcag aagaagactg ccggctggga gccctccacc
     1081 gagaagctgt acccccagga cggcgtgctg aagggcgaga tctcccacgc cctgaagctg
     1141 aaggacggcg gccactacac ctgcgacttc aagaccgtgt acaaggccaa gaagcccgtg
     1201 cagctgcccg gcaaccacta cgtggactcc aagctggaca tcaccaacca caacgaggac
     1261 tacaccgtgg tggagcagta cgagcacgcc gaggcccgcc actccggctc ccagtccgga
     1321 ctcagatctc gagctcaagc ttcgaattct gcagtcgaca tggacgaagc ggatcggcgg
     1381 ctcctgcggc ggtgccggct gcggctggtg gaagagctgc aggtggacca gctctgggac
     1441 gccctgctga gccgcgagct gttcaggccc catatgatcg aggacatcca gcgggcaggc
     1501 tctggatctc ggcgggatca ggccaggcag ctgatcatag atctggagac tcgagggagt
     1561 caggctcttc ctttgttcat ctcctgctta gaggacacag gccaggacat gctggcttcg
     1621 tttctgcgaa ctaacaggca agcagcaaag ttgtcgaagc caaccctaga aaaccttacc
     1681 ccagtggtgc tcagaccaga gattcgcaaa ccagaggttc tcagaccgga aacacccaga
     1741 ccagtggaca ttggttctgg aggatttggt gatgtcggtg ctcttgagag tttgagggga
     1801 aatgcagatt tggcttacat cctgagcatg gagccctgtg gccactgcct cattatcaac
     1861 aatgtgaact tctgccgtga gtccgggctc cgcacccgca ctggctccaa catcgactgt
     1921 gagaagttgc ggcgtcgctt ctcctcgctg catttcatgg tggaggtgaa gggcgacctg
     1981 actgccaaga aaatggtgct ggctttgctg gagctggcgc agcaggacca cggtgctctg
     2041 gactgctgcg tggtggtcat tctctctcac ggctgtcagg ccagccacct gcagttccca
     2101 ggggctgtct acggcacaga tggatgccct gtgtcggtcg agaagattgt gaacatcttc
     2161 aatgggacca gctgccccag cctgggaggg aagcccaagc tctttttcat ccaggccgcg
     2221 ggtggggagc agaaagacca tgggtttgag gtggcctcca cttcccctga agacgagtcc
     2281 cctggcagta accccgagcc agatgccacc ccgttccagg aaggtttgag gaccttcgac
     2341 cagctggacg ccatatctag tttgcccaca cccagtgaca tctttgtgtc ctactctact
     2401 ttcccaggtt ttgtttcctg gagggacccc aagagtggct cctggtacgt tgagaccctg
     2461 gacgacatct ttgagcagtg ggctcactct gaagacctgc agtccctcct gcttagggtc
     2521 gctaatgctg tttcggtgaa agggatttat aaacagatgc ctggttgctt taatttcctc
     2581 cggaaaaaac ttttctttaa aacatcataa ggccagggcc cctcaccctg ccttatcttg
     2641 caggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     2701 tacttacttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2761 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2821 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2881 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2941 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3001 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3061 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3121 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3181 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3241 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3301 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     3361 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3421 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3481 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3661 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3781 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3841 gcaaagatcg atatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     3901 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     3961 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     4021 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     4081 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     4141 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     4201 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     4261 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     4321 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     4381 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     4441 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     4501 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     4561 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     4621 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     4681 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     4741 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     4801 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     4861 ccgagggcaa aggaatagag ttctagagga tcataatcag ccataccaca tttgtagagg
     4921 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     4981 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5041 tcacaaattt cacaaataaa gcattttttt cactgcctcg cgatccacaa ccatggcgaa
     5101 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5161 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5221 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5281 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5341 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5401 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5461 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5521 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5581 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5641 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5701 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5761 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5821 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5881 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5941 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6001 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6061 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6121 ctgagatagg tgcctcactg attaagcatt ggtaaggcgc gccaggccct gccatagcct
     6181 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6241 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6301 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6361 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6421 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6481 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6541 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6601 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6661 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6721 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6781 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6841 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6901 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6961 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattct
//
