LOCUS Exported 6737 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS SIRT1.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6737) AUTHORS Hallows WC, Lee S, Denu JM TITLE Sirtuins deacetylate and activate mammalian acetyl-CoA synthetases. JOURNAL Proc Natl Acad Sci U S A. 2006 Jul 5. 103(27):10230-5. PUBMED 16790548 REFERENCE 2 (bases 1 to 6737) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6737 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 19..63 /label=T5 promoter /note="bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator" protein_bind 39..55 /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)." protein_bind 71..87 /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 106..117 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 136..153 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 2203..2297 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS 2341..3000 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2407..2426) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 3065..3151 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(3244..4326) /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 4288..4307 /label=LacI-R /note="LacI, reverse primer" promoter complement(4327..4404) /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." primer_bind complement(4488..4510) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 4596..4736 /label=bom /note="basis of mobility region from pBR322" primer_bind 4651..4670 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(4751..4768) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4922..5510) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5002..5021) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5681..6541) /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 6304..6323 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6542..6646) /gene="bla" /label=AmpR promoter primer_bind 6714..6732 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 cgtcttcacc tcgagaaatc ataaaaaatt tatttgcttt gtgagcggat aacaattata 61 atagattcaa ttgtgagcgg ataacaattt cacacagaat tcattaaaga ggagaaatta 121 actatgagag gatcgcatca ccatcaccat cacggatcca tggcggacga ggcggcggcg 181 gcaggcgggg agcaagaggc ccaggcgact gcggcggctg gggaaggaga caatgggccg 241 ggcctgcagg gcccatctcg ggagccaccg ctggccgaca acttgtacga cgaagacgac 301 gacgacgagg gcgaggagga ggaagaggcg gcggcggcgg cgattgggta ccgagataac 361 cttctgttcg gtgatgaaat tatcactaat ggttttcatt cctgtgaaag tgatgaggag 421 gatagagcct cacatgcaag ctctagtgac tggactccaa ggccacggat aggtccatat 481 acttttgttc agcaacatct tatgattggc acagatcctc gaacaattct taaagattta 541 ttgccggaaa caatacctcc acctgagttg gatgatatga cactgtggca gattgttatt 601 aatatccttt cagaaccacc aaaaaggaaa aaaagaaaag atattaatac aattgaagat 661 gctgtgaaat tactgcaaga gtgcaaaaaa attatagttc taactggagc tggggtgtct 721 gtttcatgtg gaatacctga cttcaggtca agggatggta tttatgctcg ccttgctgta 781 gacttcccag atcttccaga tcctcaagcg atgtttgata ttgaatattt cagaaaagat 841 ccaagaccat tcttcaagtt tgcaaaggaa atatatcctg gacaattcca gccatctctc 901 tgtcacaaat tcatagcctt gtcagataag gaaggaaaac tacttcgcaa ctatacccag 961 aacatagaca cgctggaaca ggttgcggga atccaaagga taattcagtg tcatggttcc 1021 tttgcaacag catcttgcct gatttgtaaa tacaaagttg actgtgaagc tgtacgagga 1081 gatattttta atcaggtagt tcctcgatgt cctaggtgcc cagctgatga accgcttgct 1141 atcatgaaac cagagattgt gttttttggt gaaaatttac cagaacagtt tcatagagcc 1201 atgaagtatg acaaagatga agttgacctc ctcattgtta ttgggtcttc cctcaaagta 1261 agaccagtag cactaattcc aagttccata ccccatgaag tgcctcagat attaattaat 1321 agagaacctt tgcctcatct gcattttgat gtagagcttc ttggagactg tgatgtcata 1381 attaatgaat tgtgtcatag gttaggtggt gaatatgcca aactttgctg taaccctgta 1441 aagctttcag aaattactga aaaacctcca cgaacacaaa aagaattggc ttatttgtca 1501 gagttgccac ccacacctct tcatgtttca gaagactcaa gttcaccaga aagaacttca 1561 ccaccagatt cttcagtgat tgtcacactt ttagaccaag cagctaagag taatgatgat 1621 ttagatgtgt ctgaatcaaa aggttgtatg gaagaaaaac cacaggaagt acaaacttct 1681 aggaatgttg aaagtattgc tgaacagatg gaaaatccgg atttgaagaa tgttggttct 1741 agtactgggg agaaaaatga aagaacttca gtggctggaa cagtgagaaa atgctggcct 1801 aatagagtgg caaaggagca gattagtagg cggcttgatg gtaatcagta tctgtttttg 1861 ccaccaaatc gttacatttt ccatggcgct gaggtatatt cagactctga agatgacgtc 1921 ttatcctcta gttcttgtgg cagtaacagt gatagtggga catgccagag tccaagttta 1981 gaagaaccca tggaggatga aagtgaaatt gaagaattct acaatggctt agaagatgag 2041 cctgatgttc cagagagagc tggaggagct ggatttggga ctgatggaga tgatcaagag 2101 gcaattaatg aagctatatc tgtgaaacag gaagtaacag acatgaacta tccatcaaac 2161 aaatcatgtc gacctgcagc caagcttaat tagctgagct tggactcctg ttgatagatc 2221 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt 2281 tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa ggaagctaaa 2341 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa 2401 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat 2461 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt 2521 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt 2581 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa 2641 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat 2701 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag 2761 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg 2821 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc 2881 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat 2941 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa 3001 tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag 3061 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt 3121 gtcggtgaac gctctcctga gtaggacaaa tccgccctct agattacgtg cagtcgatga 3181 taagctgtca aacatgagaa ttgtgcctaa tgagtgagct aacttacatt aattgcgttg 3241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 3301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag 3361 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg 3421 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat 3481 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg 3541 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag 3601 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat 3661 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt 3721 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc 3781 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg 3841 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac 3901 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat 3961 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc 4021 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat 4081 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag 4141 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc 4201 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac 4261 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg 4321 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa 4381 ggttttgcac cattcgatgg tgtcggaatt tcgggcagcg ttgggtcctg gccacgggtg 4441 cgcatgatct agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg 4501 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt 4561 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc 4621 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 4681 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct 4741 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4801 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4861 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4921 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4981 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5041 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5101 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5161 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 5221 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5281 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5341 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 5401 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 5461 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 5521 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 5581 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 5641 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 5701 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 5761 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 5821 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5881 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5941 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6001 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6061 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6121 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6181 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6241 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6301 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6361 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6421 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 6481 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 6541 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 6601 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 6661 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 6721 gtatcacgag gcccttt //