LOCUS Exported 8927 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CD95-1-2xPP7, NLS-PCP-DNCR2-P2a-BFP. ACCESSION . VERSION . KEYWORDS GWF303 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8927 /organism="synthetic DNA construct" /mol_type="other DNA" 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 2063..2096 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." 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(4270..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 8077..8110 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." 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 8838..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 //