LOCUS       sequence-8476-ex        4623 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 4623)
  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 4623)
  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::caspase-3 (p17) [TU#814]
FEATURES             Location/Qualifiers
     source          1..4623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2063..2085)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2185..2204
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2282..2300)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2368..2472
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2473..2541,2542..3333)
                     /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(2691..2710)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3504..4092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3993..4012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4246..4263
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4380..4401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4416..4421,4422..4439,4440..4446)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4454..4470
                     /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     4459..4481
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4478..4494
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4478..4494
                     /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 tccaaaaatg gctagcgcac
      421 agctggagaa gaaactgcag gctctggaga agaaactggc acagctggag tggaaaaacc
      481 aggcactgga gaagaaactg gcacagggtg gaagcggtcc ccgggattgg ccaaaggacc
      541 caaaggtatg tttcgaatga tactaacata acatagaaca ttttcaggag gacccttgga
      601 gggtaccaat ggagaacact gaaaactcag tggattcaaa atccattaaa aatttggaac
      661 caaagatcat acatggaagc gaatcaatgg actctggaat atccctggac aacagttata
      721 aaatggatta tcctgagatg ggtttatgta taataattaa taataagaat tttcataaaa
      781 gcactggaat gacatctcgg tctggtacag atgtcgatgc agcaaacctc agggaaacat
      841 tcagaaactt gaaatatgaa gtcaggaata aaaatgatct tacacgtgaa gaaattgtgg
      901 aattgatgcg tgatgtttct aaagaagatc acagcaaaag gagcagtttt gtttgtgtgc
      961 ttctgagcca tggtgaagaa ggaataattt ttggaacaaa tggacctgtt gacctgaaaa
     1021 aaataacaaa ctttttcaga ggggatcgtt gtagaagtct aactggaaaa cccaaacttt
     1081 tcattattca ggcctgccgt ggtacagaac tggactgtgg cattgagaca gactgatcta
     1141 gaattccaac tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag
     1201 gggccgctgt catcagagta agtttaaact gagttctact aactaacgag taatatttaa
     1261 attttcagca tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc
     1321 tacatgctct ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaaacttct
     1381 tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa attgtgtaga
     1441 ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc ctccccccat
     1501 taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt atgttttttt
     1561 tacttctgat aaattttttt tgaaacatca tagaaaaaac cgcacacaaa ataccttatc
     1621 atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct gggcctctca
     1681 tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat ttttcaatca
     1741 agtgaaagtt tatgaaatta attttcctgc ttttgctttt tgggggtttc ccctattgtt
     1801 tgtcaagagt ttcgaggacg gcgtttttct tgctaaaatc acaagtattg atgagcacga
     1861 tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga gtagaagttg
     1921 ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag aatacattcc
     1981 caatatacca aacataactg tttcctacta gtcggccgta cgggcccttt cgtctcgcgc
     2041 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2101 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2161 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2221 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cggccttaag
     2281 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2341 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2401 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2461 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2521 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2581 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2641 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2701 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2761 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2821 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2881 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2941 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3001 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3061 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3121 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3181 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3241 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3301 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3361 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3421 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     3901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4141 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4201 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4261 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4321 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4381 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     4441 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     4501 tacgccaagc tgtaagttta aacatgatct tactaactaa ctattctcat ttaaattttc
     4561 agagcttaaa aatggctgaa atcactcaca acgatggata cgctaacaac ttggaaatga
     4621 aat
//
