LOCUS Exported 6135 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of Flag tagged human FOXO3 in mammalian cells. ACCESSION . VERSION . KEYWORDS Flag-FOXO3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6135) AUTHORS Moparthi L, Koch S TITLE A uniform expression library for the exploration of FOX transcription factor biology JOURNAL Elsevier 2020 REFERENCE 2 (bases 1 to 6135) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6135 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..53 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 35..52 /label=SP6 /note="SP6 promoter, forward primer" regulatory 85..94 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 94..117 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(2165..2182) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2187..2321 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2211..2230) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2265..2284 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2434..2454) /label=T3 /note="T3 promoter, forward primer" promoter complement(2434..2452) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2473..2489) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2473..2489) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2486..2508) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2497..2513 /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(2521..2551) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2566..2587 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2704..2721) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2875..3463) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2955..2974) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3634..4494) /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 4257..4276 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4495..4599) /gene="bla" /label=AmpR promoter rep_origin complement(4625..5080) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4762..4783) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4974..4993 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5158..6135 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag 61 ctacttgttc tttttgcagg atcagccacc atggattaca aggacgacga cgataaggga 121 tccgcagagg caccggcttc cccggccccg ctctctccgc tcgaagtgga gctggacccg 181 gagttcgagc cccagagccg tccgcgatcc tgtacgtggc ccctgcaaag gccggagctc 241 caagcgagcc ctgccaagcc ctcgggggag acggccgccg actccatgat ccccgaggag 301 gaggacgatg aagacgacga ggacggcggg ggacgggccg gctcggccat ggcgatcggc 361 ggcggcggcg ggagcggcac gctgggctcc gggctgctcc ttgaggactc ggcccgggtg 421 ctggcacccg gagggcaaga ccccgggtct gggccagcca ccgcggcggg cgggctgagc 481 gggggtacac aggcgctgct gcagcctcag caaccgctgc caccgccgca gccgggggcg 541 gctgggggct ccgggcagcc gaggaaatgt tcgtcgcggc ggaacgcctg gggaaacctg 601 tcctacgcgg acctgatcac ccgcgccatc gagagctccc cggacaaacg gctcactctg 661 tcccagatct acgagtggat ggtgcgttgc gtgccctact tcaaggataa gggcgacagc 721 aacagctctg ccggctggaa gaactccatc cggcacaacc tgtcactgca tagtcgattc 781 atgcgggtcc agaatgaggg aactggcaag agctcttggt ggatcatcaa ccctgatggg 841 gggaagagcg gaaaagcccc ccggcggcgg gctgtctcca tggacaatag caacaagtat 901 accaagagcc gtggccgcgc agccaagaag aaggcagccc tgcagacagc ccccgaatca 961 gctgacgaca gtccctccca gctctccaag tggcctggca gccccacgtc acgcagcagt 1021 gatgagctgg atgcgtggac ggacttccgt tcacgcacca attctaacgc cagcacagtc 1081 agtggccgcc tgtcgcccat catggcaagc acagagttgg atgaagtcca ggacgatgat 1141 gcgcctctct cgcccatgct ctacagcagc tcagccagcc tgtcaccttc agtaagcaag 1201 ccgtgcacgg tggaactgcc acggctgact gatatggcag gcaccatgaa tctgaatgat 1261 gggctgactg aaaacctcat ggacgacctg ctggataaca tcacgctccc gccatcccag 1321 ccatcgccca ctgggggact catgcagcgg agctctagct tcccgtatac caccaagggc 1381 tcgggcctgg gctccccaac cagctccttt aacagcacgg tgttcggacc ttcatctctg 1441 aactccctac gccagtctcc catgcagacc atccaagaga acaagccagc taccttctct 1501 tccatgtcac actatggtaa ccagacactc caggacctgc tcacttcgga ctcacttagc 1561 cacagcgatg tcatgatgac acagtcggac cccttgatgt ctcaggccag caccgctgtg 1621 tctgcccaga attcccgccg gaacgtgatg cttcgcaatg atccgatgat gtcctttgct 1681 gcccagccta accagggaag tttggtcaat cagaacttgc tccaccacca gcaccaaacc 1741 cagggcgctc ttggtggcag ccgtgccttg tcgaattctg tcagcaacat gggcttgagt 1801 gagtccagca gccttgggtc agccaaacac cagcagcagt ctcctgtcag ccagtctatg 1861 caaaccctct cggactctct ctcaggctcc tccttgtact caactagtgc aaacctgccc 1921 gtcatgggcc atgagaagtt ccccagcgac ttggacctgg acatgttcaa tgggagcttg 1981 gaatgtgaca tggagtccat tatccgtagt gaactcatgg atgctgatgg gttggatttt 2041 aactttgatt ccctcatctc cacacagaat gttgttggtt tgaacgtggg gaacttcact 2101 ggtgctaagc aggcctcatc tcagagctgg gtgccaggct gaaggatcac tcgagcctct 2161 agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg 2221 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta 2281 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc 2341 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc 2401 caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt 2461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 2521 caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 2581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 2641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 2701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 2761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 2821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 2881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 2941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 3001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 3061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 3121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 3181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 3241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 3301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 3361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 3421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 3481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 3541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 3601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 3661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 3721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 3781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 3841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 3901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 3961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 4021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 4081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 4141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 4201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 4261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 4321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 4381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 4441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 4501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 4561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 4621 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc 4681 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc 4741 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac 4801 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca 4861 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg 4921 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag 4981 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc 5041 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg 5101 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc 5161 aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc 5221 aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc 5281 aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc 5341 aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc 5401 caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc 5461 aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt 5521 tcaatactat gtattggccc tatgccatat agtattccat atatgggttt tcctattgac 5581 gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc 5641 ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc 5701 atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat 5761 ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg 5821 gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa 5881 atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat 5941 tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc 6001 cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg 6061 ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat 6121 gctcgtttag ggaac //