LOCUS Exported 5714 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCDNA3 Flag DLC1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5714) AUTHORS Lei K, Davis RJ TITLE JNK phosphorylation of Bim-related members of the Bcl2 family induces Bax-dependent apoptosis. JOURNAL Proc Natl Acad Sci U S A. 2003 Mar 4. 100(5):2432-7. PUBMED 12591950 REFERENCE 2 (bases 1 to 5714) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5714 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(63..80) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(234..822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(314..333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(993..1853) /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 1616..1635 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1854..1958) /gene="bla" /label=AmpR promoter primer_bind complement(2033..2052) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2224..2603 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2604..2807 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2758..2778 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2852..2871 /label=T7 /note="T7 promoter, forward primer" promoter 2852..2870 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2899..2922 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(3256..3274) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3257..3274) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3294..3311) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3300..3524 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3570..3998 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3657..3676) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3867..3888 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4007..4027) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4012..4341 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4192..4327 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4254..4273 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4408..5202 /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(4462..4481) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5072..5091 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5376..5497 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5413..5432) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5467..5486 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5546..5562) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5546..5562) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5559..5581) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5570..5586 /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(5594..5624) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5639..5660 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 2881 cttacggcag ccaacatgga ctataaggac gatgatgaca aaggatcctg cgaccgaaag 2941 gccgtgatca aaaatgcgga catgtcggaa gagatgcaac aggactcggt ggagtgcgct 3001 actcaggcgc tggagaaata caacatagag aaggacattg cggctcatat caagaaggaa 3061 tttgacaaga agtacaatcc cacctggcat tgcatcgtgg ggaggaactt cggtagttat 3121 gtgacacatg aaaccaaaca cttcatctac ttctacctgg gccaagtggc cattcttctg 3181 ttcaaatctg gttaagaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca 3241 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac 3301 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct 3361 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct 3421 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg 3481 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag 3541 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc 3601 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 3661 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 3721 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 3781 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 3841 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 3901 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 3961 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag 4021 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 4081 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 4141 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc 4201 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 4261 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt 4321 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat 4381 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct 4441 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc 4501 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc 4561 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc 4621 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg 4681 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 4741 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 4801 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt 4861 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc 4921 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 4981 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg 5041 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag 5101 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 5161 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg 5221 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct 5281 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc 5341 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg 5401 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 5461 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct 5521 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 5581 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 5641 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 5701 tgtcgtgcca gctg //