LOCUS       sequence-424374-        6635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6635)
  AUTHORS   Stennicke HR, Deveraux QL, Humke EW, Reed JC, Dixit VM, Salvesen GS
  TITLE     Caspase-9 can be activated without proteolytic processing.
  JOURNAL   J Biol Chem. 1999 Mar 26. 274(13):8359-62.
  PUBMED    10085063
REFERENCE   2  (bases 1 to 6635)
  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: "J Biol 
            Chem. 1999 Mar 26. 274(13):8359-62."
COMMENT     Sequence Label: pcDNA3-Casp9 C287A
FEATURES             Location/Qualifiers
     source          1..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..56)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(40..56)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(53..75)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    64..80
                     /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(88..94,95..112,113..118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    133..154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(271..288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(442..1030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(522..541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1201..1992,1993..2061))
                     /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     1824..1843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2062..2166)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2241..2260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2432..2811
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2812..3015
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2966..2986
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3060..3079
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3060..3078
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(3848..3859)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(4385..4403)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4386..4403)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4423..4440)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4429..4653
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4699..5127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4786..4805)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4996..5017
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5136..5156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5141..5470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5321..5456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5383..5402
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5537..6331
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5591..5610)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6201..6220
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6505..6626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6542..6561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6596..6615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
       61 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
      121 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
      181 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      241 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      301 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      361 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      421 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      481 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      541 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      601 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      661 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      721 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      781 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      841 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      901 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1021 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1081 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1141 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1201 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1261 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1321 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     1381 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1441 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1501 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1561 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1621 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1681 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1741 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1801 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1861 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1921 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1981 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2041 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2101 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2161 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     2221 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     2281 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     2341 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     2401 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     2461 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2521 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2581 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2641 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2701 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     2761 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     2821 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     2881 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     2941 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3001 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     3061 aatacgactc actataggga gacccaagct tgcggccgcg aattcatcga tagatcccat
     3121 ggacgaagcg gatcggcggc tcctgcggcg gtgccggctg cggctggtgg aagagctgca
     3181 ggtggaccag ctctgggacg ccctgctgag ccgcgagctg ttcaggcccc atatgatcga
     3241 ggacatccag cgggcaggct ctggatctcg gcgggatcag gccaggcagc tgatcataga
     3301 tctggagact cgagggagtc aggctcttcc tttgttcatc tcctgcttag aggacacagg
     3361 ccaggacatg ctggcttcgt ttctgcgaac taacaggcaa gcagcaaagt tgtcgaagcc
     3421 aaccctagaa aaccttaccc cagtggtgct cagaccagag attcgcaaac cagaggttct
     3481 cagaccggaa acacccagac cagtggacat tggttctgga ggatttggtg atgtcggtgc
     3541 tcttgagagt ttgaggggaa atgcagattt ggcttacatc ctgagcatgg agccctgtgg
     3601 ccactgcctc attatcaaca atgtgaactt ctgccgtgag tccgggctcc gcacccgcac
     3661 tggctccaac atcgactgtg agaagttgcg gcgtcgcttc tcctcgctgc atttcatggt
     3721 ggaggtgaag ggcgacctga ctgccaagaa aatggtgctg gctttgctgg agctggcgca
     3781 gcaggaccac ggtgctctgg actgctgcgt ggtggtcatt ctctctcacg gctgtcaggc
     3841 cagccacctg cagttcccag gggctgtcta cggcacagat ggatgccctg tgtcggtcga
     3901 gaagattgtg aacatcttca atgggaccag ctgccccagc ctgggaggga agcccaagct
     3961 ctttttcatc caggccgctg gtggggagca gaaagaccat gggtttgagg tggcctccac
     4021 ttcccctgaa gacgagtccc ctggcagtaa ccccgagcca gatgccaccc cgttccagga
     4081 aggtttgagg accttcgacc agctggacgc catatctagt ttgcccacac ccagtgacat
     4141 ctttgtgtcc tactctactt tcccaggttt tgtttcctgg agggacccca agagtggctc
     4201 ctggtacgtt gagaccctgg acgacatctt tgagcagtgg gctcactctg aagacctgca
     4261 gtccctcctg cttagggtcg ctaatgctgt ttcggtgaaa gggatttata aacagatgcc
     4321 tggttgcttt aatttcctcc ggaaaaaact tttctttaaa acatcataat ctagagggcc
     4381 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     4441 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4501 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4561 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4621 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4681 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4741 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4801 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4861 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4921 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4981 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5041 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5101 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     5161 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5221 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5281 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5341 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5401 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5461 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     5521 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5581 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5641 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5701 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5761 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5821 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5881 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5941 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     6001 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     6061 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     6121 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     6181 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     6241 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     6301 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6361 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6421 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6481 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6541 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6601 ttgtccaaac tcatcaatgt atcttatcat gtctg
//
