LOCUS       sequence-384887-        6186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible Expression of N-Terminal 3X Flag Mouse C325A Caspase-9 
            Minus Linker + Small Subunit Domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6186)
  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 6186)
  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-LD...
FEATURES             Location/Qualifiers
     source          1..6186
                     /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(2194..2211)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2200..2424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2470..2898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2557..2576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2767..2788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2907..2927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2912..3241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3092..3227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3154..3173
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3289..3336
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3355..3729
                     /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    3859..3980
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3896..3915)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3950..3969
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4029..4045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4029..4045)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4042..4064)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4053..4069
                     /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(4077..4083,4084..4101,4102..4107))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4122..4143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4260..4277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4431..5019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4511..4530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5190..5981,5982..6050))
                     /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     5813..5832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6051..6155)
                     /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 gatggacgag gcggaccggc
     1081 agctcctgcg gcgatgcagg gtgcgcctag tgagcgagct gcaagtcgcg gagctctggg
     1141 acgctctgct gagtcgagag ctcttcacgc gcgacatgat cgaggatatt cagcaggcag
     1201 gatctgggtc tcggcgggat caggccaggc agctggtcac agaccttgag acccgaggga
     1261 ggcaggccct tcctctcttc atctcctgct tagaggacac aggccaaggc accctggctt
     1321 cactcttgca aagcggtcgg caagcagcca agcaggatcc agaggctgtt aaacccctag
     1381 accacctggt gcctgtggtc ctgggaccaa tgggactcac agcaaaggag cagagagtag
     1441 tgaagctgga cccgtcacag cctgccgtgg gaaacctcac cccagtggtg ctggggccag
     1501 aagagctctg gcctgctcgg ctcaagccag aggttctcag accagaaaca cccaggcccg
     1561 tggacattgg ttctggcgga gctcatgatg tctgtgttcc agggaagatc aggggacatg
     1621 cagatatggc atacaccctg gattcggatc cctgtggcca ctgcctcatc atcaacaatg
     1681 tgaacttctg cccttcctcg gggctcggca cacgcacggg ctccaacttg gaccgtgaca
     1741 aacttgagca ccgattccgc tggctgcgct tcatggtgga ggtgaagaac gacctgactg
     1801 ccaagaaaat ggtcacggct ttgatggaga tggcacaccg gaaccaccgt gccctggact
     1861 gctttgtggt ggtcatcctc tctcatggct gccaggccag ccacctccag ttcccgggtg
     1921 ctgtctatgg gacagatgga tgctccgtgt ccattgagaa aattgtgaat atcttcaacg
     1981 ggagcggctg ccccagcctg ggagggaagc ccaagctctt cttcatccag gccgcgggtg
     2041 gtgagcagaa agaccatggc tttgaggtgg cctgcacttc ctctcaaggc aggaccttgg
     2101 acagtgactc tgagccagat tgagaattct gcagatatcc agcacagtgg cggccgctcg
     2161 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     2221 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2281 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2341 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2401 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2461 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2521 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2581 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2641 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2701 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2761 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2821 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2881 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2941 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3001 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3061 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3121 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3181 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3241 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     3301 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     3361 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     3421 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     3481 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     3541 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     3601 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     3661 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     3721 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3781 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     3841 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3901 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3961 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4021 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4081 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4141 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4201 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4261 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4321 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4381 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4441 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4501 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4561 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4621 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4681 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4741 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4801 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4861 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4921 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4981 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5041 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5101 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5161 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5221 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5281 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5341 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5401 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5461 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5521 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5581 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5641 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5701 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5761 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5821 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5881 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5941 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6001 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6061 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6121 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6181 gacgtc
//
