LOCUS Exported 6540 bp ds-DNA circular SYN 19-JAN-2022 DEFINITION Mammalian expression of human SIRT2 with flag tag. ACCESSION . VERSION . KEYWORDS SIRT2 Flag SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6540) AUTHORS North BJ, Marshall BL, Borra MT, Denu JM, Verdin E TITLE The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase. JOURNAL Mol Cell. 2003 Feb . 11(2):437-44. PUBMED 12620231 REFERENCE 2 (bases 1 to 6540) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 19, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6540 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1132..1155 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(1190..1207) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1196..1420 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1466..1894 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1553..1572) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1763..1784 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1903..1923) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1908..2237 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2088..2223 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2150..2169 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2304..3098 /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(2358..2377) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2968..2987 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3272..3393 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3309..3328) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3363..3382 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3442..3458) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3442..3458) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3455..3477) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3466..3482 /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(3490..3520) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3535..3556 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3673..3690) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3844..4432) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3924..3943) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4603..5463) /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 5226..5245 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5464..5568) /gene="bla" /label=AmpR promoter primer_bind complement(5643..5662) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 5834..6213 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6214..6417 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6368..6388 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 6462..6481 /label=T7 /note="T7 promoter, forward primer" promoter 6462..6480 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 atggcagagc agaccgactc agattcagac tctgagggag gagccgctgg tggagaagca 61 gacatggact tcctgcggaa cttattctcc cagacgctca gcctgggcag ccagaaggag 121 cgtctgctgg acgagctgac cttggaaggg gtggcccggt acatgcagag cgaacgctgt 181 cgcagagtca tctgtttggt gggagctgga atctccacat ccgcaggcat ccccgacttt 241 cgctctccat ccaccggcct ctatgacaac ctagagaagt accatcttcc ctacccagag 301 gccatctttg agatcagcta tttcaagaaa catccggaac ccttcttcgc cctcgccaag 361 gaactctatc ctgggcagtt caagccaacc atctgtcact acttcatgcg cctgctgaag 421 gacaaggggc tactcctgcg ctgctacacg cagaacatag ataccctgga gcgaatagcc 481 gggctggaac aggaggactt ggtggaggcg cacggcacct tctacacatc acactgcgtc 541 agcgccagct gccggcacga atacccgcta agctggatga aagagaagat cttctctgag 601 gtgacgccca agtgtgaaga ctgtcagagc ctggtgaagc ctgatatcgt cttttttggt 661 gagagcctcc cagcgcgttt cttctcctgt atgcagtcag acttcctgaa ggtggacctc 721 ctcctggtca tgggtacctc cttgcaggtg cagccctttg cctccctcat cagcaaggca 781 cccctctcca cccctcgcct gctcatcaac aaggagaaag ctggccagtc ggaccctttc 841 ctggggatga ttatgggcct cggaggaggc atggactttg actccaagaa ggcctacagg 901 gacgtggcct ggctgggtga atgcgaccag ggctgcctgg cccttgctga gctccttgga 961 tggaagaagg agctggagga ccttgtccgg agggagcacg ccagcataga tgcccagtcg 1021 ggggcggggg tccccaaccc cagcacttca gcttccccca agaagtcccc gccacctgcc 1081 aaggacgagg ccaggacaac agagagggag aaaccccagg aattcggagg agactacaag 1141 gacgacgatg acaagtaatc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg 1201 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 1261 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 1321 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 1381 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 1441 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 1501 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 1561 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 1621 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 1681 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 1741 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 1801 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 1861 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 1921 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 1981 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 2041 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 2101 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 2161 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 2221 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 2281 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 2341 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 2401 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 2461 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 2521 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 2581 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 2641 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 2701 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 2761 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 2821 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 2881 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 2941 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 3001 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 3061 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 3121 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 3181 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 3241 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 3301 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 3361 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 3421 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 3481 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 3541 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 3601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 3661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 3721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 3781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 3841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 3901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 3961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 4141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 4201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 4261 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 4321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 4381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 4441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 4501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 4561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 4621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 4681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 4741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 4801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 4861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 4921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 4981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 5101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 5161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 5221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 5281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 5341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 5401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 5461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 5521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 5581 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc 5641 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg 5701 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac 5761 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc 5821 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca 5881 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 5941 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 6001 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 6061 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 6121 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 6181 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 6241 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 6301 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 6361 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc 6421 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg 6481 gagacccaag ctggctagcg tttaaactta agcttggtac cgagctcgga tccgaattct //