LOCUS       sequence-8473-ex        5307 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 5307)
  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 5307)
  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 ced-3 (p15)::nz [TU#806]
FEATURES             Location/Qualifiers
     source          1..5307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1786..1797
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(2747..2769)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2869..2888
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2966..2984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3052..3156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3157..3225,3226..4017)
                     /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(3375..3394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4188..4776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4677..4696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4930..4947
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5064..5085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5100..5105,5106..5123,5124..5130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5138..5154
                     /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     5143..5165
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5162..5178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5162..5178
                     /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 tccaaaatgg gagttcctgc
      421 atttcttcgt cgtggatggg acaatcgaga cgggccattg ttcaattttc ttggatgtgt
      481 gcggccgcaa gttcaggttg caatttaatt tcttgaatga gaatattcct tcaaaaaatc
      541 taaaatagat ttttattcca gaaagtccga cgaaaaaatt gcgatataat tacgaaattt
      601 gtgataaaat gacaaaccaa tcagcatcgt cgatctccgc ccacttcatc ggattggttt
      661 gaaagtgggc ggagtgaatt gctgattggt cgcagttttc agtttagagg gaatttaaaa
      721 atcgcctttt cgaaaattaa aaattgattt tttcaatttt ttcgaaaaat attccgatta
      781 ttttatattc tttggagcga aagccccgtc ctgtaaacat ttttaaatga taattaataa
      841 atttttgcag caagtgtgga gaaagaagcc gagccaagct gacattctga ttgcatacgc
      901 aacgacagct caatatgttt cgtggagaaa cagtgctcgt ggatcatggt tcattcaagc
      961 cgtctgtgaa gtgttctcga cacacgcaaa ggatatggat gttgttgagc tgctgactga
     1021 agtcaataag aaggtcgctt gtggatttca gacatcacag ggatcgaata ttttgaaaca
     1081 gatgccagag gtacttgaac aacaatcgat ctgctaactt tttaaggcac agaaaaatag
     1141 gcagagcgtc cttttgcaag cctgccgcgc gtcaacctag aattttagtt tttagctaaa
     1201 atgattgatt ttgaatattt tatgctaatt tttttgcgtt aaattttgaa atagtcacta
     1261 tttatcgggt ttccagtaaa aaatgtttat tagccattgg attttactga aaacgaaaat
     1321 ttgtagtttt tcaacgaaat ttatcgattt ttaaatgtaa aaaaaaatag cgaaaataca
     1381 tcaacatcaa gcatttaagc caaaaattgt taactcattt aaaaaattaa ttcaaagttg
     1441 tccacgagta ttacacggtt ggcgcgcggc aagtttgcaa aacgacgctc cgcctctttt
     1501 tctgtgcggc ttgaaaacaa gggtatcggt tttagatttt tccccaaaat ttaaattaaa
     1561 tttcagatga catcccgcct gctcaaaaag ttctactttt ggccggaggc acgaaactct
     1621 gccgtccccc gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac
     1681 atagaacatt ttcaggagga cccttggagg gtaccaggct ctggctctgg cgctcagctt
     1741 aagaaagagc tgcaggcaaa caagaaagag ctggctcagc tgaagtggga actgcaggca
     1801 ctgaagaaag aactggctca gtgagaattc caactgagcg ccggtcgcta ccattaccaa
     1861 cttgtctggt gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc
     1921 tactaactaa cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta
     1981 agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt
     2041 tttgttatta tcaaaaaaac ttcttcttaa tttctttgtt ttttagcttc ttttaagtca
     2101 cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa
     2161 aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc
     2221 tgtgtacact tcttatgttt tttttacttc tgataaattt tttttgaaac atcatagaaa
     2281 aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat
     2341 ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg
     2401 agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc
     2461 tttttggggg tttcccctat tgtttgtcaa gagtttcgag gacggcgttt ttcttgctaa
     2521 aatcacaagt attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc
     2581 tcagtggaag gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt
     2641 gccttaaatg acagaataca ttcccaatat accaaacata actgtttcct actagtcggc
     2701 cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2761 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2821 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     2881 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2941 aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc
     3001 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3061 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4321 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4561 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4861 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4921 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4981 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5041 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5101 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5161 acaggaaaca gctatgacca tgattacgcc aagctgtaag tttaaacatg atcttactaa
     5221 ctaactattc tcatttaaat tttcagagct taaaaatggc tgaaatcact cacaacgatg
     5281 gatacgctaa caacttggaa atgaaat
//
