LOCUS       sequence-8478-ex        3242 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 3242)
  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 3242)
  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 cz::caspase-3 (p17) [TU#818]
FEATURES             Location/Qualifiers
     source          1..3242
                     /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"
     regulatory      350..359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1156..1237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1193..1212)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1248..1265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1419..2007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1499..1518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2178..2969,2970..3038))
                     /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     2801..2820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3039..3143)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3211..3229
                     /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 ccaccatggc
      361 tagcgcacag ctggagaaga aactgcaggc tctggagaag aaactggcac agctggagtg
      421 gaaaaaccag gcactggaga agaaactggc acagggtgga agcggtcccc gggattggcc
      481 aaaggaccca aaggaggacc cttggagggt accaatggag aacactgaaa actcagtgga
      541 ttcaaaatcc attaaaaatt tggaaccaaa gatcatacat ggaagcgaat caatggactc
      601 tggaatatcc ctggacaaca gttataaaat ggattatcct gagatgggtt tatgtataat
      661 aattaataat aagaattttc ataaaagcac tggaatgaca tctcggtctg gtacagatgt
      721 cgatgcagca aacctcaggg aaacattcag aaacttgaaa tatgaagtca ggaataaaaa
      781 tgatcttaca cgtgaagaaa ttgtggaatt gatgcgtgat gtttctaaag aagatcacag
      841 caaaaggagc agttttgttt gtgtgcttct gagccatggt gaagaaggaa taatttttgg
      901 aacaaatgga cctgttgacc tgaaaaaaat aacaaacttt ttcagagggg atcgttgtag
      961 aagtctaact ggaaaaccca aacttttcat tattcaggcc tgccgtggta cagaactgga
     1021 ctgtggcatt gagacagact gatctagagg atcataatca gccataccac atttgtagag
     1081 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1141 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1201 atcacaaatt tcacaaataa agcatttttt tcactgcctc gagcttcctc gctcactgac
     1261 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1321 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1381 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1441 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1501 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1561 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     1621 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1681 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1741 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1801 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     1861 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1921 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1981 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2041 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2101 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2161 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2221 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2281 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2341 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2401 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2461 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2521 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2581 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2641 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2701 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2761 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2821 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2881 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2941 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3001 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3061 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3121 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     3181 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     3241 ca
//
