LOCUS       sequence-384891-        6000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible Expression of N-Terminal 3X Flag Mouse C325A Caspase-9 
            Mutant Minus CARD Domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6000)
  AUTHORS   Han J, Hou W, Goldstein LA, Stolz DB, Watkins SC, Rabinowich H
  TITLE     A Complex between Atg7 and Caspase-9: A NOVEL MECHANISM OF 
            CROSS-REGULATION BETWEEN AUTOPHAGY AND APOPTOSIS.
  JOURNAL   J Biol Chem. 2014 Mar 7;289(10):6485-6497. doi: 
            10.1074/jbc.M113.536854. Epub 2013 Dec 20.
  PUBMED    24362031
REFERENCE   2  (bases 1 to 6000)
  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: 
            "10.1074/jbc.M113.536854"; journalName: "J Biol Chem"; date: 
            "2014-03-7- 7"; volume: "289"; issue: "10"; pages: "6485-6497"
COMMENT     Sequence Label: Tet-inducible 3xFlag Mouse C325A Caspase-9 
            Delta-CARD
FEATURES             Location/Qualifiers
     source          1..6000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             996..1061
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2008..2025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2014..2238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2284..2712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2371..2390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2581..2602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2721..2741)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2726..3055
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2906..3041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2968..2987
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3103..3150
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3169..3543
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3673..3794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3710..3729)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3764..3783
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3843..3859)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3843..3859)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3856..3878)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3867..3883
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3891..3897,3898..3915,3916..3921))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3936..3957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4074..4091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4245..4833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4325..4344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5004..5795,5796..5864))
                     /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     5627..5646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5865..5969)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg ccatggacta caaagaccat gacggtgatt
     1021 ataaagatca tgacatcgat tacaaggatg acgatgacaa ggtctgtgtt ccagggaaga
     1081 tcaggggaca tgcagatatg gcatacaccc tggattcgga tccctgtggc cactgcctca
     1141 tcatcaacaa tgtgaacttc tgcccttcct cggggctcgg cacacgcacg ggctccaact
     1201 tggaccgtga caaacttgag caccgattcc gctggctgcg cttcatggtg gaggtgaaga
     1261 acgacctgac tgccaagaaa atggtcacgg ctttgatgga gatggcacac cggaaccacc
     1321 gtgccctgga ctgctttgtg gtggtcatcc tctctcatgg ctgccaggcc agccacctcc
     1381 agttcccggg tgctgtctat gggacagatg gatgctccgt gtccattgag aaaattgtga
     1441 atatcttcaa cgggagcggc tgccccagcc tgggagggaa gcccaagctc ttcttcatcc
     1501 aggccgcggg tggtgagcag aaagaccatg gctttgaggt ggcctgcact tcctctcaag
     1561 gcaggacctt ggacagtgac tctgagccag atgctgtccc ctatcaggaa ggcccaaggc
     1621 ccttggacca gctggatgct gtgtcaagtt tgcctacccc cagtgacatc cttgtgtcct
     1681 actccacctt cccaggtttt gtctcctgga gggacaagaa aagtggctcc tggtacatcg
     1741 agaccttgga tggcattctg gagcagtggg ctcgctctga agacctgcag tccctccttc
     1801 tcagggttgc caatgctgtt tctgcgaaag ggacttacaa gcagattcct ggctgtttta
     1861 acttcctccg gaaaaagctg ttttttaaaa cttcatgagg ctcggggctg ctctctgcct
     1921 cattattctc tccgaaggaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta
     1981 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2041 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2101 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2161 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2221 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     2281 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2341 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2401 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2461 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2521 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2581 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2641 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2701 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2761 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2821 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2881 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2941 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3001 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3061 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     3121 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
     3181 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     3241 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     3301 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     3361 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     3421 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     3481 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     3541 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3601 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3661 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3721 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3781 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     3841 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3901 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3961 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4021 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4081 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4141 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4201 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4261 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4321 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4381 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4441 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4501 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4561 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4621 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4681 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4741 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4801 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4861 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4921 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4981 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5041 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5101 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5161 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5221 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5281 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5341 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5401 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5461 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5521 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5581 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5641 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5701 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5761 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5821 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5881 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5941 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
