LOCUS       sequence-8475-ex        4399 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 4399)
  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 4399)
  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: Pmec-18 caspase-3 (p12)::nz [TU#813]
FEATURES             Location/Qualifiers
     source          1..4399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             878..889
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1839..1861)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1961..1980
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2058..2076)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2144..2248
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2249..2317,2318..3109)
                     /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     complement(2467..2486)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3280..3868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3769..3788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4022..4039
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4156..4177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4192..4197,4198..4215,4216..4222)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4230..4246
                     /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)."
     primer_bind     4235..4257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4254..4270
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4254..4270
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttcaat taattcgtct actatccacg tgtcgattcg aagacagttc acatgagtca
       61 ttgactttga tctggaacat attttttaat atagaaattt ccgattttga ttacctttgc
      121 catgcacact tgcactttgc aacaataacc atttgagcga ttcgctcgag tagttctcgc
      181 gctcttcttg acatcagatg aaaataacaa aaagaggaga gcattgacca aatgtatcaa
      241 gattgcatgg taaaacatag caaatgtttg tcatccgatg ggttggctcc ttgttgcact
      301 ggtatgcatt tgtgatgcga aacaggagct cttccagaag cagtcttgca cttcttgtat
      361 tcattccttt gcctcgcctt ccaagaaggt tgtgagcgga tccaccgcca tgagtggtgt
      421 tgatgatgac atggcgtgtc ataaaatacc agtggaggcc gacttcttgt atgcatactc
      481 cacagcacct ggttattatt cttggcgaaa ttcaaaggat ggctcctggt tcatccagtc
      541 gctttgtgcc atgctgaaac agtatgccga caagcttgaa tttatgcaca ttcttacccg
      601 ggttaaccga aaggtggcaa cagaatttga gtccttttcc tttgacgcta cttttcatgc
      661 aaagaaacag attccatgta ttgtttccat gctcacaaaa gaactctatt tttatcaccc
      721 ccgggattgg ccaaaggacc caaaggtatg tttcgaatga tactaacata acatagaaca
      781 ttttcaggag gacccttgga gggtaccagg ctctggctct ggcgctcagc ttaagaaaga
      841 gctgcaggca aacaagaaag agctggctca gctgaagtgg gaactgcagg cactgaagaa
      901 agaactggct cagtgagaat tccaactgag cgccggtcgc taccattacc aacttgtctg
      961 gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt taaactgagt tctactaact
     1021 aacgagtaat atttaaattt tcagcatctc gcgcccgtgc ctctgacttc taagtccaat
     1081 tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta tttttgttat
     1141 tatcaaaaaa acttcttctt aatttctttg ttttttagct tcttttaagt cacctctaac
     1201 aatgaaattg tgtagattca aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg
     1261 tgctccctcc ccccattaat aataattcta tcccaaaatc tacacaatgt tctgtgtaca
     1321 cttcttatgt tttttttact tctgataaat tttttttgaa acatcataga aaaaaccgca
     1381 cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt atttcttcgc
     1441 acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt ggagtatttt
     1501 tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt gctttttggg
     1561 ggtttcccct attgtttgtc aagagtttcg aggacggcgt ttttcttgct aaaatcacaa
     1621 gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga
     1681 aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa
     1741 tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg gccgtacggg
     1801 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1861 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1921 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     1981 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     2041 atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     2101 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2161 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2221 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2281 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2341 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2401 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2461 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2521 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2581 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2641 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2701 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2761 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2821 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2881 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2941 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3001 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3061 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3121 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3181 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3361 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3421 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3481 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3541 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3601 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3661 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3721 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3781 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3841 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3901 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3961 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4021 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4081 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4141 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     4201 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     4261 cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat
     4321 tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct
     4381 aacaacttgg aaatgaaat
//
