LOCUS       sequence-8477-ex        3020 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 3020)
  AUTHORS   Chelur DS, Chalfie M
  TITLE     Targeted cell killing by reconstituted caspases.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Feb 13. 104(7):2283-8.
  PUBMED    17283333
REFERENCE   2  (bases 1 to 3020)
  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: "Proc Natl 
            Acad Sci U S A. 2007 Feb 13. 104(7):2283-8."
COMMENT     Sequence Label: pTRE-Tight caspase-3 (p12)::nz [TU#817]
FEATURES             Location/Qualifiers
     source          1..3020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..318
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    119..137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    155..173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    190..208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     291..315
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             782..793
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    934..1015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(971..990)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1026..1043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1197..1785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1277..1296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1956..2747,2748..2816))
                     /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     2579..2598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2817..2921)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2989..3007
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
       61 tagagaacga tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc
      121 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
      181 tcgagtttat ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
      241 gagaacgtat gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta
      301 gtgaaccgtc agatcgcctg gagaattcga gctcggtacc cggggatccg ccaccatgag
      361 tggtgttgat gatgacatgg cgtgtcataa aataccagtg gaggccgact tcttgtatgc
      421 atactccaca gcacctggtt attattcttg gcgaaattca aaggatggct cctggttcat
      481 ccagtcgctt tgtgccatgc tgaaacagta tgccgacaag cttgaattta tgcacattct
      541 tacccgggtt aaccgaaagg tggcaacaga atttgagtcc ttttcctttg acgctacttt
      601 tcatgcaaag aaacagattc catgtattgt ttccatgctc acaaaagaac tctattttta
      661 tcacccccgg gattggccaa aggacccaaa ggaggaccct tggagggtac caggctctgg
      721 ctctggcgct cagcttaaga aagagctgca ggcaaacaag aaagagctgg ctcagctgaa
      781 gtgggaactg caggcactga agaaagaact ggctcagtga tctagaggat cataatcagc
      841 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
      901 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
      961 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcctcga
     1021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1381 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1621 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     2581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2941 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     3001 acgaggccct ttcgtcttca
//
