LOCUS       sequence-261566-        8927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CD95-1-2xPP7, NLS-PCP-DNCR2-P2a-BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8927)
  AUTHORS   Foight GW, Wang Z, Wei CT, Jr Greisen P, Warner KM, 
            Cunningham-Bryant D, Park K, Brunette TJ, Sheffler W, Baker D, Maly 
            DJ
  TITLE     Multi-input chemical control of protein dimerization for programming
            graded cellular responses.
  JOURNAL   Nat Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8.
  PUBMED    31501561
REFERENCE   2  (bases 1 to 8927)
  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: "Nat 
            Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8."
COMMENT     Sequence Label: GWF303
FEATURES             Location/Qualifiers
     source          1..8927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             151..171
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1246..1275
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1285..1341
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1342..2040
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     protein_bind    complement(2063..2096)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2152..2740
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2205..2225)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2623..2634)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2743..2759)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2744..2760)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3269..3449
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3511..4099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3591..3610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4270..5061,5062..5130))
                     /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     4893..4912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5131..5235)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5310..5329)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5501..5880
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5882..6080
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6098..6278
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6325..6450
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6947..7180
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7365..7409
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7707..7824
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(8077..8110)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     8114..8134
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     terminator      8383..8402
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     enhancer        8534..8837
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8838..8927,1..114)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
       61 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      121 gtgaaccgtc agatccgcta gcccaccatg cccaaaaaga aaagaaaagt gggtagtatg
      181 tccaaaacca tcgttctttc ggtcggcgag gctactcgca ctctgactga gatccagtcc
      241 accgcagacc gtcagatctt cgaagagaag gtcgggcctc tggtgggtcg gctgcgcctc
      301 acggcttcgc tccgtcaaaa cggagccaag accgcgtatc gcgtcaacct aaaactggat
      361 caggcggacg tcgttgattc cggacttccg aaagtgcgct acactcaggt atggtcgcac
      421 gacgtgacaa tcgttgcgaa tagcaccgag gcctcgcgca aatcgttgta cgatttgacc
      481 aagtccctcg tcgcgacctc gcaggtcgaa gatcttgtcg tcaaccttgt gccgctgggc
      541 cgtggctccg gctctggttc ttccgacgaa gaagaagcgc gcgagctgat cgagcgtgcg
      601 aaggaggctg cggaacgcgc gcaagaagct gctgagcgta ccggtgatcc gcgcgtgcgt
      661 gaactcgctc gtgagctgaa acgcctggct caggaagctg ctgaagaagt gaagcgtgac
      721 ccgtcttcct ccgatgtgaa cgaagcgctc aaactcatcg tggaagctat cgaggcggct
      781 gtggacgcgc tcgaagcggc tgaacgcacc ggcgacccgg aggttcgtga gctggcgcgt
      841 gagctcgtgc gcctcgcggt tgaggctgcg gaggaggtgc aacgtaaccc atcctcctct
      901 gacgtgaatg aggctctcca cagcatagtt tacgctattg aagctgccat ttttgcattg
      961 gaagctgccg aaaggactgg tgacccggaa gtgagagaac tcgcccgaga gcttgtaaga
     1021 ctggcggttg aggcggctga agaagtccag cgaaatccgt ccagccggaa tgtcgagcac
     1081 gcactgatga ggattgtgtt ggccatttac ctggcagaag aaaatctccg cgaagctgag
     1141 gagtctggcg atccggaaaa acgtgaaaag gctcgcgaac gtgttcgtga ggcggtggaa
     1201 cgtgcggaag aggttcagcg tgatccatcc ggttggctga atcatgaaca aaaacttatt
     1261 tccgaagagg atctgggaag cggagctact aacttcagcc tgctgaagca ggctggagac
     1321 gtggaggaga accctggacc tagcgagctg attaaggaga acatgcacat gaagctgtac
     1381 atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc
     1441 tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc
     1501 ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag
     1561 ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc
     1621 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc
     1681 ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag
     1741 aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg
     1801 gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc
     1861 aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat
     1921 gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac
     1981 gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaattga
     2041 taagaattcg tcgagggacc taataacttc gtatagcata cattatacga agttatacat
     2101 gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct
     2161 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2221 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2281 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2341 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2401 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2461 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2521 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     2581 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2641 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2701 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     2761 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     2821 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     2881 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     2941 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     3001 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     3061 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     3121 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     3181 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     3241 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     3301 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     3361 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3421 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
     3481 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3541 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3601 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3661 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3721 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3781 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3841 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3901 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3961 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4021 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4081 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4141 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4201 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4261 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4321 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4381 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4441 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4501 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4561 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4621 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4681 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4741 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4801 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4861 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4921 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4981 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5041 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5101 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5221 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     5281 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5341 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     5401 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     5461 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     5521 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     5581 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     5641 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     5701 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     5761 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     5821 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     5881 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     5941 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     6001 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     6061 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
     6121 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6181 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6241 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     6301 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     6361 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     6421 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     6481 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     6541 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     6601 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     6661 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     6721 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     6781 cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     6841 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     6901 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     6961 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     7021 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     7081 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     7141 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     7201 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     7261 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     7321 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     7381 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     7441 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     7501 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     7561 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     7621 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     7681 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     7741 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     7801 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     7861 ttagtaccgg gcccgctcta gagatccgac gcgccatctc taggcccgcg ccggccccct
     7921 cgcacggact tgtgggagaa gctcggctac tcccctgccc cggttaattt gcatataata
     7981 tttcctagta actatagagg cttaatgtgc gataaaagac agataatctg ttctttttaa
     8041 tactagctac attttacatg ataggcttgg atttctataa cttcgtatag catacattat
     8101 acgaagttat aaacagcaca aaaggaaact caccctaact gtaaagtaat tgtgtgtttt
     8161 gagactataa gtatcccttg gagaaccacc ttgttggtac agcagaagcc tttagaagtt
     8221 taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt tatcaacttg
     8281 aaaaagtggc accgagtcgg tgcgggagct aaggagttta tatggaaacc cttagcctgc
     8341 tgcgtaagga gtttatatgg aaacccttac gcagcagttc cctttttttg ttttttatgt
     8401 ctctgcagag ttcggtctcg agtactagga tccattaggc ggccgcgtgg ataaccgtat
     8461 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     8521 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     8581 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     8641 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     8701 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     8761 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     8821 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     8881 ttgactcacg gggatttcca agtctccacc ccattgacgt caatggg
//
