LOCUS Exported 7737 bp ds-DNA circular SYN 04-FEB-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMSCV-F-del Casp9.IRES.GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7737) AUTHORS Straathof KC, Pule MA, Yotnda P, Dotti G, Vanin EF, Brenner MK, Heslop HE, Spencer DM, Rooney CM TITLE An inducible caspase 9 safety switch for T-cell therapy. JOURNAL Blood. 2005 Jun 1. 105(11):4247-54. PUBMED 15728125 REFERENCE 2 (bases 1 to 7737) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 4, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7737 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 579..920 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 987..1403 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1333..1355 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1371..1387 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1427..1747 /codon_start=1 /product="F36V mutant of FK506-binding protein FKBP12" /label=DmrB /note="binds synthetic ligands that do not bind wild-type FKBP" /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" CDS complement(2099..2110) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" CDS 2624..2650 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2841..2858) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3068..3087 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 3249..3968 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3249..3965 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3294..3315) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3555..3574) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3902..3923 /label=EGFP-C /note="EGFP, forward primer" LTR 4130..4644 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 4806..4822 /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)." promoter complement(4830..4860) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4875..4896 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5013..5030) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5184..5772) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5264..5283) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5943..6803) /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 6566..6585 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6804..6908) /gene="bla" /label=AmpR promoter primer_bind 6976..6994 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7032..7054) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7154..7173 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7367..7389 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7517..7536 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 7736..515 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc 301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata 421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca 481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga 541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg 601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt 661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa 721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga 781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag 841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa 901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg 961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct 1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga 1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag 1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc 1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc 1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg 1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag 1381 ccctcactcc ttctctaggc gccggaatta cgccaccatg ctcgagggag tgcaggtgga 1441 gactatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca 1501 ctacaccggg atgcttgaag atggaaagaa agttgattcc tcccgggaca gaaacaagcc 1561 ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca 1621 gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac 1681 tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa 1741 actggaatct ggcggtggat ccggagtcga cggatttggt gatgtcggtg ctcttgagag 1801 tttgagggga aatgcagatt tggcttacat cctgagcatg gagccctgtg gccactgcct 1861 cattatcaac aatgtgaact tctgccgtga gtccgggctc cgcacccgca ctggctccaa 1921 catcgactgt gagaagttgc ggcgtcgctt ctcctcgctg catttcatgg tggaggtgaa 1981 gggcgacctg actgccaaga aaatggtgct ggctttgctg gagctggcgc ggcaggacca 2041 cggtgctctg gactgctgcg tggtggtcat tctctctcac ggctgtcagg ccagccacct 2101 gcagttccca ggggctgtct acggcacaga tggatgccct gtgtcggtcg agaagattgt 2161 gaacatcttc aatgggacca gctgccccag cctgggaggg aagcccaagc tctttttcat 2221 ccaggcctgt ggtggggagc agaaagacca tgggtttgag gtggcctcca cttcccctga 2281 agacgagtcc cctggcagta accccgagcc agatgccacc ccgttccagg aaggtttgag 2341 gaccttcgac cagctggacg ccatatctag tttgcccaca cccagtgaca tctttgtgtc 2401 ctactctact ttcccaggtt ttgtttcctg gagggacccc aagagtggct cctggtacgt 2461 tgagaccctg gacgacatct ttgagcagtg ggctcactct gaagacctgc agtccctcct 2521 gcttagggtc gctaatgctg tttcggtgaa agggatttat aaacagatgc ctggttgctt 2581 taatttcctc cggaaaaaac ttttctttaa aacatcagtc gactatccgt acgacgtacc 2641 agactacgca ctcgactaag aattcatcga gcgggatcaa ttccgccccc cccctaacgt 2701 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac 2761 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag 2821 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa 2881 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag 2941 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga 3001 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag 3061 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc 3121 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag 3181 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg 3241 ataataccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg 3301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg 3361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct 3421 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc 3481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca 3541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg 3601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc 3661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc 3721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc 3781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg 3841 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc 3901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt 3961 acaagtaaag cggccgcgac tctagagtcg acctgcaggc atgcaagctt caggtagccg 4021 gctaacgtta acaaccggta cctctagaac tatagctagc atgcgcaaat ttaaagcgct 4081 gatatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc 4141 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca 4201 taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc 4261 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc 4321 cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc 4381 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct 4441 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc 4501 gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg 4561 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac 4621 tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct 4681 gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca 4741 tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg 4801 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 4861 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 4921 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 4981 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5041 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5101 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5161 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5221 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5281 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5341 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 5401 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5461 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5521 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5581 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 5641 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5701 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 5761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 5821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 5881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 5941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 6841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 6901 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg 6961 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat 7021 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg 7081 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc 7141 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 7201 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg 7261 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 7321 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 7381 tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg 7441 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag 7501 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc 7561 cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca 7621 ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag 7681 agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaa //