LOCUS Exported 6906 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-Casp8 C360A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6906) AUTHORS Stennicke HR, Salvesen GS TITLE Biochemical characteristics of caspases-3, -6, -7, and -8. JOURNAL J Biol Chem. 1997 Oct 10. 272(41):25719-23. PUBMED 9325297 REFERENCE 2 (bases 1 to 6906) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6906 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..48) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 275..295 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin complement(304..732) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(414..435) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 626..645 /label=F1ori-R /note="F1 origin, reverse primer" polyA_signal 778..1002 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 991..1008 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" promoter 1028..1046 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1028..1045 /label=SP6 /note="SP6 promoter, forward primer" CDS complement(1141..1167) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2623..2642) /label=T7 /note="T7 promoter, forward primer" promoter complement(2624..2642) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(2687..2890) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(2716..2736) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 2891..3270 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 3442..3461 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 3536..3640 /gene="bla" /label=AmpR promoter CDS 3641..4501 /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(3859..3878) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4672..5260 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5161..5180 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5414..5431 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5548..5569 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5584..5614 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5622..5638 /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)." primer_bind 5627..5649 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5646..5662 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5646..5662 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 5711..5832 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5722..5741) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5776..5795 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(6006..6800) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6117..6136) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 6727..6746 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(6867..290) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6881..110 /label=SV40 ori /note="SV40 origin of replication" ORIGIN 1 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt 61 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt 121 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct 181 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata 241 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga 301 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg 361 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt 421 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct 481 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt 541 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa 601 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc 661 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc 721 tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca 781 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt 841 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt 901 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg 961 gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcgag 1021 ctctagcatt taggtgacac tatagaatag ggccctctag atgcatgctc gagcggccgc 1081 cagtgtgatg gatatctgca gaattccagc acactggcgg ccgttactag tggatcctca 1141 agcgtagtct ggcacgtcgt atgggtaatc agaagggaag acaagttttt ttcttagtgt 1201 gaaagtaggc tgaggcatct gtttccccat gtttttcttg tcatccttgt tgcttacttc 1261 atagttcact tcagtcagga tggtgagaat atcatcgcct cgaggacatc gctctctcag 1321 gctctggcaa agtgactgga tgtaccaggt tccctctgca gggtttcggt aggaaacaca 1381 gttattcaca gtggccatcc ccagcagaaa gtcagcctca tccgggatat atctcgtttg 1441 aggtgatgat aaatccattt ctaaataggg ttgctcctct gaatcagtct caacaggtat 1501 acctttctgg tagttatccc cctgagcagc ctgaataaaa aacactttgg gttttccagc 1561 aagggaaggg cacttcaaac cagtgaactg agatgtcagc tcatagatgg gggcctcctg 1621 tccatcagtg ccatagatga tgcccttgtc tccatgggag aggatacagc agatgaagca 1681 gtccatgtta ctgtggtcca tgagttggta gattttcaaa atctcataga tttgctctac 1741 tgtgcagtca tcgtggggct tgatctcaaa atgaagctct tcaaaggtcg tggtcaaagc 1801 ccctgcatcc aagtgtgttc cattcctgtc cctaatgctg tgaagtttgg gcactttctc 1861 ccgtgctttt gcaaaattgt gattgttgat gatcagacag tatccccgag gtttgctttt 1921 catttggtaa actttgtcca aagtctgtga ttcactatcc tgttctcttg gagagtccga 1981 gattgtcatt accccacaca actcctcccc atttgaaaat tcatcaggac ttccttcaag 2041 gctgctgctt ctctctttgc tgaattcttc atagtcgttg attatcttca gcaggctctt 2101 gttgatttgg gcacagactc ttttcaggat gtccaacttt ccttctccca ggatgaccct 2161 cttctccatc tctatgaaaa tatccagcag gttcatgtca tcatccagtt tgcatttgga 2221 gatttcctct tgcaaaagaa acttaaaaga cctcaattct gatctgctca cttcttctga 2281 aatctgatag agcatgaccc tgtaggcaga aatttgagcc ctgcctggtg tctgaagttc 2341 cctttccatc tcctcctttc tagtgtttag gtaggtaatc agcaaatcca gtctattaat 2401 tcggaagagc agctccttca ggaaggacag attgctttcc tccaacattc tcttttcctg 2461 gagtctctgg aataacatca aggcatcctt gatgggttct tgcttccttt gcggaatgta 2521 gtccaggctc aggaacttga gggaggccag atcttcactg tccagttgtt ccccaatatc 2581 ataaagattt ctgctgaagt ccatggtacc aagcttgggt ctccctatag tgagtcgtat 2641 taatttcgat aagccagtaa gcagtgggtt ctctagttag ccagagagct ctgcttatat 2701 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac 2761 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt 2821 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa 2881 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt 2941 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa 3001 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta 3061 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg 3121 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg 3181 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt 3241 actattaata actagtcaat aatcaatgtc aacgcgtata tctggcccgt acatcgcgaa 3301 gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca attgtcggtc aagccttgcc 3361 ttgttgtagc ttaaattttg ctcgcgcact actcagcgac ctccaacaca caagcaggga 3421 gcagatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccataggg 3481 gatcgggaga tctcccgatc cgtcgacgtc aggtggcact tttcggggaa atgtgcgcgg 3541 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 3601 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 3661 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 3721 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 3781 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 3841 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga 3901 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 3961 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 4021 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 4081 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 4141 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac 4201 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 4261 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 4321 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 4381 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 4441 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 4501 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 4561 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 4621 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 4681 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 4741 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 4801 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc 4861 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 4921 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 4981 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 5041 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 5101 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 5161 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 5221 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 5281 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 5341 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 5401 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc 5461 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg 5521 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca 5581 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt 5641 tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg tcgacggtat 5701 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa 5761 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc 5821 aataaacaag ttggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat 5881 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga 5941 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt 6001 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg 6061 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata 6121 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg 6181 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg 6241 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct 6301 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc 6361 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga 6421 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca 6481 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc 6541 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat 6601 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa 6661 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc 6721 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc 6781 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatccgaaa 6841 atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa aaagcctcct 6901 cactac //