LOCUS Exported 6194 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses caspase 3 with C163S to prevent autocatalysis in bacterial cells. ACCESSION . VERSION . KEYWORDS pET21b-Caspase-3(C163S) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6194) AUTHORS Pop C, Chen YR, Smith B, Bose K, Bobay B, Tripathy A, Franzen S, Clark AC TITLE Removal of the pro-domain does not affect the conformation of the procaspase-3 dimer. JOURNAL Biochemistry. 2001 Nov 27;40(47):14224-35. PUBMED 11714276 REFERENCE 2 (bases 1 to 6194) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6194 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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)." RBS 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 926..943 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(996..1014) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1010..1057 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1094..1549 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1181..1200) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1391..1412 /label=F1ori-F /note="F1 origin, forward primer" promoter 1575..1679 /gene="bla" /label=AmpR promoter CDS 1680..2540 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1898..1917) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2711..3299 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3200..3219 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3453..3470 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3485..3624 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3550..3569) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3710..3732 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3726..3917) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4726..5808) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 5770..5789 /label=LacI-R /note="LacI, reverse primer" promoter complement(5809..5886) /gene="lacI" /label=lacI promoter primer_bind 6092..6111 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atatacatat ggagaacact gaaaactcag tggattcaaa 121 atccattaaa aatttggaac caaagatcat acatggaagc gaatcaatgg actctggaat 181 atccctggac aacagttata aaatggatta tcctgagatg ggtttatgta taataattaa 241 taataagaat tttcataaaa gcactggaat gacatctcgg tctggtacag atgtcgatgc 301 agcaaacctc agggaaacat tcagaaactt gaaatatgaa gtcaggaata aaaatgatct 361 tacacgtgaa gaaattgtgg aattgatgcg tgatgtttct aaagaagatc acagcaaaag 421 gagcagtttt gtttgtgtgc ttctgagcca tggtgaagaa ggaataattt ttggaacaaa 481 tggacctgtt gacctgaaaa aaataacaaa ctttttcaga ggggatcgtt gtagaagtct 541 aactggaaaa cccaaacttt tcattattca ggcctcccgc ggtacagaac tggactgtgg 601 cattgagaca gacagtggtg ttgatgatga catggcgtgt cataaaatac cagtggaggc 661 cgacttcttg tatgcatact ccacagcacc tggttattat tcttggcgaa attcaaagga 721 tggctcctgg ttcatccagt cgctttgtgc catgctgaaa cagtatgccg acaagcttga 781 atttatgcac attcttaccc gggttaaccg aaaggtggca acagaatttg agtccttttc 841 ctttgacgct acttttcatg caaagaaaca gattccatgt attgtttcca tgctcacaaa 901 agaactctat ttttatcacc tcgagcacca ccaccaccac cactgagatc cggctgctaa 961 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc 1021 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg 1081 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 1141 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 1201 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 1261 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 1321 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 1381 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 1441 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 1501 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt 1561 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 1621 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta 1681 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg 1741 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac 1801 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg 1861 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc 1921 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg 1981 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat 2041 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg 2101 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg 2161 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc 2221 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt 2281 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct 2341 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc 2401 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca 2461 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct 2521 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt 2581 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 2641 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 2701 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 2761 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 2821 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag 2881 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 2941 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 3001 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 3061 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 3121 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 3181 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 3241 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 3301 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 3361 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg 3421 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag 3481 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg 3541 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg 3601 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga 3661 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc 3721 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca 3781 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg 3841 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt 3901 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg 3961 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg 4021 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa 4081 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc 4141 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt 4201 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac 4261 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca 4321 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc 4381 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga 4441 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg 4501 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc 4561 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg 4621 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat 4681 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc 4741 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 4801 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg 4861 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc 4921 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc 4981 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat 5041 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat 5101 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc 5161 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg 5221 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa 5281 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt 5341 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc 5401 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg 5461 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga 5521 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga 5581 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc 5641 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt 5701 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata 5761 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct 5821 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat 5881 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta 5941 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc 6001 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga 6061 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa 6121 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct 6181 cgatcccgcg aaat //