LOCUS       sequence-8474-ex        5525 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 5525)
  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 5525)
  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 cz::ced-3 (p17) [TU#807]
FEATURES             Location/Qualifiers
     source          1..5525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2965..2987)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3087..3106
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3184..3202)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3270..3374
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3375..3443,3444..4235)
                     /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(3593..3612)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4406..4994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4895..4914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5148..5165
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5282..5303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5318..5323,5324..5341,5342..5348)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5356..5372
                     /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     5361..5383
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5380..5396
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5380..5396
                     /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 tgtgagcatg ggatccaaaa atggctagcg
      421 cacagctgga gaagaaactg caggctctgg agaagaaact ggcacagctg gagtggaaaa
      481 accaggcact ggagaagaaa ctggcacagg gtggaagcgg tccccgggat tggccaaagg
      541 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
      601 ggagggtacc ggtagcacca accataagcc gtgttttcga cgagaaaacc atgtacagaa
      661 acttctcgag tcctcgtgga atgtgcctca tcataaataa tgaacacttt gagcagatgc
      721 caacacggaa tggtaccaag gccgacaagg acaatcttac caatttgttc agatgcatgg
      781 gctatacggt tatttgcaag gacaatctga cgggaagggt acggcgaaat tatattaccc
      841 aaacgcgaaa tttttgccat ttttgcgccg aaaatgtggc gcccggtctc gacacgacaa
      901 tttgtgttaa atgcaaaaat gtataatttt gcaaaaaaca aaattttgaa cttccgcgaa
      961 aatgatttac ctagtttcga aattttcgtt ttttccggct acattatgtg ttttttctta
     1021 gtttttctat aatatttgat gtaaaaaacc gtttgtaaat tttcagacaa ttttccgcat
     1081 acaaaacttg atagcacgaa atcaattttc tgaattttca aaattatcca aaaatgcaca
     1141 atttaaaatt tgtgaaaatt ggcaaacggt gtttcaatat gaaatgtatt tttaaaaact
     1201 ttaaaaacca ctccggaaaa gcaataaaaa tcaaaacaac gtcacaattc aaattcaaaa
     1261 gttattcatc cgatttgttt atttttgcaa aatttgaaaa aatcatgaag gatttagaaa
     1321 agttttataa cattttttct agatttttca aaattttttt taacaaatcg agaaaaagag
     1381 aatgaaaaat cgattttaaa aatatccaca gcaacgagag tttgaaatta cagtactcct
     1441 taaaggcgca caccccattt gcattgacca aaaatttgtc gtgtcgagac caggtaccgt
     1501 agtttttgtc gcaaaaattg caccattgga caataaacct tcctaatcac caaaaagtaa
     1561 aattgaaatc ttcgaaaagc caaaaaattc aaaaaaaaag tcgaatttcg attttttttt
     1621 tggttttttg gtcccaaaaa ccaaaaaatc aatttttcgt caaaatacca aaagaaaccc
     1681 aaaaaatttc ccagccttgt tcctaatgta aactgatatt taatttccag ggaatgctcc
     1741 tgacaattcg agactttgcc aaacacgaat cacacggaga ttctgcgata ctcgtgattc
     1801 tatcacacgg agaagagaat gtgattattg gagttgatga tataccgatt agtacacacg
     1861 agatatatga tcttctcaac gcggcaaatg ctccccgtct ggcgaataag ccgaaaatcg
     1921 tttttgtgca ggcttgtcga ggcggttcgt tttttatttt aattttaata taaatatttt
     1981 aaataaattc attttcagaa cgtcgtgaca atggattccc agtcttggat tctgtcgact
     2041 gagaattcca actgagcgcc ggtcgctacc attaccaact tgtctggtgt caaaaataat
     2101 aggggccgct gtcatcagag taagtttaaa ctgagttcta ctaactaacg agtaatattt
     2161 aaattttcag catctcgcgc ccgtgcctct gacttctaag tccaattact cttcaacatc
     2221 cctacatgct ctttctccct gtgctcccac cccctatttt tgttattatc aaaaaaactt
     2281 cttcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta
     2341 gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc
     2401 attaataata attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt
     2461 tttacttctg ataaattttt tttgaaacat catagaaaaa accgcacaca aaatacctta
     2521 tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt ctgggcctct
     2581 catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga atttttcaat
     2641 caagtgaaag tttatgaaat taattttcct gcttttgctt tttgggggtt tcccctattg
     2701 tttgtcaaga gtttcgagga cggcgttttt cttgctaaaa tcacaagtat tgatgagcac
     2761 gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt gagtagaagt
     2821 tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac agaatacatt
     2881 cccaatatac caaacataac tgtttcctac tagtcggccg tacgggccct ttcgtctcgc
     2941 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3001 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3061 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     3121 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta
     3181 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3241 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3301 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3361 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3421 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3481 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3541 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3601 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3661 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3721 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3781 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3841 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3901 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3961 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4021 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4081 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4141 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4201 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4261 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4321 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4381 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4441 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4501 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4561 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4621 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4681 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4741 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     4801 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4861 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4921 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4981 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5041 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5101 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5161 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5221 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5281 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5341 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5401 attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc atttaaattt
     5461 tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca acttggaaat
     5521 gaaat
//
