LOCUS       sequence-214299-        6798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a tandem caspase-9 dimer, governed by a StaPL(AI) module, 
            in mammalian cells. [AI = asunaprevir inhibited]..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6798)
  AUTHORS   Jacobs CL, Badiee RK, Lin MZ
  TITLE     StaPLs: versatile genetically encoded modules for engineering 
            drug-inducible proteins.
  JOURNAL   Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z.
            Epub 2018 Jul 2.
  PUBMED    29967496
REFERENCE   2  (bases 1 to 6798)
  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"; doi: 
            "10.1038/s41592-018-0041-z"; journalName: "Nat Methods"; date: 
            "2018-07"; volume: "15"; issue: "7"; pages: "523-526"
COMMENT     Sequence Label: pCS6-StaPLd-Casp9
FEATURES             Location/Qualifiers
     source          1..6798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(157..225,226..1017)
                     /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(375..394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    1119..1152
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      1230..1818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1719..1738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1972..1989
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(2063..2167)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        2277..2580
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2581..2784
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2735..2755
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2781..2805
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     2910..2926
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2910..2926
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2937..2955
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2937..2954
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    2956..2980
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2999..3008
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3326..3337)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3872..3898
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4943..4954)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(5478..5502)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5503..5522)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5504..5522)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5533..5550)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5533..5549)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5542..5564)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    5954..6088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5991..6010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6045..6064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6214..6668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6301..6320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6510..6531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gctggaggga gaataaaggt ctaagatgtg cgatagaggg aagtcgcatt gaattatgtg
       61 ctgtgtaggg atcgctggta tcaaatatgt gtgcccaccc ctggcatgag acaataaccc
      121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
      301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
      361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
      421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
      481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
      541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
      601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
      661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
      721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
      781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
      841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
      901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
      961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1081 aggatctagg tgaagatcct ttttgataat ctcatgccat aacttcgtat aatgtatgct
     1141 atacgaagtt atggcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1861 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1921 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1981 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2041 attcattaat gcagagcttg caattcgcgc tttttcaata ttattgaagc atttatcagg
     2101 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     2161 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     2221 cattaaccta taaaaatagg cgtattacga ggccctttca cgcattagat gcatgtcgtt
     2281 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     2341 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     2401 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2461 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2521 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2581 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2641 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2701 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2761 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     2821 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcc
     2881 taggccgcgg gacggataac aatttcacac aggaaacagc tatgaccatt aggcctattt
     2941 aggtgacact atagaacaag tttgtacaaa aaagcaggct ggtaccggtc cggaattcgc
     3001 caccatggat gtcggtgctc ttgagagttt gaggggaaat gcagatttgg cttacatcct
     3061 gagcatggag ccctgtggcc actgcctcat tatcaacaat gtgaacttct gccgtgagtc
     3121 cgggctccgc acccgcactg gctccaacat cgactgtgag aagttgcggc gtcgcttctc
     3181 ctcgctgcat ttcatggtgg aggtgaaggg cgacctgact gccaagaaaa tggtgctggc
     3241 tttgctggag ctggcgcggc aggaccacgg tgctctggac tgctgcgtgg tggtcattct
     3301 ctctcacggc tgtcaggcca gccacctgca gttcccaggg gctgtctacg gcacagatgg
     3361 atgccctgtg tcggtcgaga agattgtgaa catcttcaat gggaccagct gccccagcct
     3421 gggagggaag cccaagctct ttttcatcca ggcctgtggt ggggagcaga aagaccatgg
     3481 gtttgaggtg gcctccactt cccctgaaga cgagtcccct ggcagtaacc ccgagccaga
     3541 tgccaccccg ttccaggaag gtttgaggac cttcgaccag ctggacgcca tatctagttt
     3601 gcccacaccc agtgacatct ttgtgtccta ctctactttc ccaggttttg tttcctggag
     3661 ggaccccaag agtggctcct ggtacgttga gaccctggac gacatctttg agcagtgggc
     3721 tcactctgaa gacctgcagt ccctcctgct tagggtcgct aatgctgttt cggtgaaagg
     3781 gatttataaa cagatgcctg gttgctttaa tttcctccgg aaaaaacttt tctttaaaac
     3841 atcagggggc agtggcggga gcgggcgcgc ctatccctac gatgtgcccg attacgctgg
     3901 tggatccggc gggtcagatg agatggaaga gtgctctcag cacggcggat ctggaggttc
     3961 tacaggctgc gtggtcatag tgggcaggat cgtcttgtcc ggatccggca ctagtgcgcc
     4021 catcacggcg tacgcccagc agacgagagg cctcctaggg tgtataatca ccagcctgac
     4081 tggccgggac aaaaaccaag tggagggtga ggtccagatc atgtcaactg ctacccaaac
     4141 cttcctggca acgtgcatca atggggtatg ctgggcagtc taccacgggg ccggaacgag
     4201 gaccatcgca tcacccaagg gtcctgtcat ccagatgtat accaatgtgg accaagacct
     4261 tgtgggctgg cccgctcctc aaggttcccg ctcattgaca ccctgtacct gcggctcctc
     4321 ggacctttac ctggtcacga ggcacgccga tgtcattccc gtgcgccggc gaggtgatgg
     4381 caggggtagc ctgctttcgc cccggcccat ttcctacttg aaaggctcct cggggggtcc
     4441 gctgttgtgc cccgcgggac acgccgtggg cctattcagg gccgcggtgt gcacccgtgg
     4501 agtggctaaa gcggtggact ttatccctgt ggagaaccta gagacaacca tgagatcccc
     4561 ggtgttcacg gacaactcct ctccaccagc agtcaccctg acgcacgggg gttcaggcgg
     4621 tagtgatgtc ggtgctcttg agagtttgag gggaaatgca gatttggctt acatcctgag
     4681 catggagccc tgtggccact gcctcattat caacaatgtg aacttctgcc gtgagtccgg
     4741 gctccgcacc cgcactggct ccaacatcga ctgtgagaag ttgcggcgtc gcttctcctc
     4801 gctgcatttc atggtggagg tgaagggcga cctgactgcc aagaaaatgg tgctggcttt
     4861 gctggagctg gcgcggcagg accacggtgc tctggactgc tgcgtggtgg tcattctctc
     4921 tcacggctgt caggccagcc acctgcagtt cccaggggct gtctacggca cagatggatg
     4981 ccctgtgtcg gtcgagaaga ttgtgaacat cttcaatggg accagctgcc ccagcctggg
     5041 agggaagccc aagctctttt tcatccaggc ctgtggtggg gagcagaaag accatgggtt
     5101 tgaggtggcc tccacttccc ctgaagacga gtcccctggc agtaaccccg agccagatgc
     5161 caccccgttc caggaaggtt tgaggacctt cgaccagctg gacgccatat ctagtttgcc
     5221 cacacccagt gacatctttg tgtcctactc tactttccca ggttttgttt cctggaggga
     5281 ccccaagagt ggctcctggt acgttgagac cctggacgac atctttgagc agtgggctca
     5341 ctctgaagac ctgcagtccc tcctgcttag ggtcgctaat gctgtttcgg tgaaagggat
     5401 ttataaacag atgcctggtt gctttaattt cctccggaaa aaacttttct ttaaaacatc
     5461 atagaagctt tacgcgtacc cagctttctt gtacaaagtg gtccctatag tgagtcgtat
     5521 tataagctag gcactggccg tcgttttaca acgtcgtgac tgggaaaact gctagcttgg
     5581 gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac tacctacaga
     5641 gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta aactagctgc
     5701 atatgcttgc tgcttgagag ttttgcttac tgagtatgat ttatgaaaat attatacaca
     5761 ggagctagtg attctaattg tttgtgtatt ttagattcac agtcccaagg ctcatttcag
     5821 gcccctcagt cctcacagtc tgttcatgat cataatcagc cataccacat ttgtagaggt
     5881 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     5941 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6061 catcaatgta tcttatcatg tctggatcga tcctgcatta atgaatcggc caacgcgcgg
     6121 ggagaggcgg tttgcgtatt ggctggcgta atagcgaaga ggcccgcacc gatcgccctt
     6181 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     6241 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6301 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6361 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6421 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6481 ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact
     6541 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     6601 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     6661 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6721 taaatacatt caaatatgta tccgctcatg ccaggtcttg gactggtgag aacggcttgc
     6781 tcggcagctt cgatgtgt
//
