LOCUS Exported 7524 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression construct of mouse SIRT1 short isoform (lacking Exon2). ACCESSION . VERSION . KEYWORDS pcDNA 3.1 SIRT1-deltaExon2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7524) AUTHORS Deota S, Chattopadhyay T, Ramachandran D, Armstrong E, Camacho B, Maniyadath B, Fulzele A, Gonzalez-de-Peredo A, Denu JM, Kolthur-Seetharam U TITLE Identification of a Tissue-Restricted Isoform of SIRT1 Defines a Regulatory Domain that Encodes Specificity. JOURNAL Cell Rep. 2017 Mar 28;18(13):3069-3077. doi: 10.1016/j.celrep.2017.03.012. PUBMED 28355560 REFERENCE 2 (bases 1 to 7524) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7524 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 736..745 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2794..2823 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2839..2856 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2879..2896) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2885..3109 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3155..3583 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3242..3261) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3452..3473 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3592..3612) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3597..3926 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3777..3912 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3839..3858 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3993..4787 /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(4047..4066) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4657..4676 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4963..5084 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5000..5019) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5054..5073 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5133..5149) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5133..5149) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5146..5168) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5157..5173 /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(5181..5211) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5226..5247 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5364..5381) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5535..6123) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5615..5634) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6294..7154) /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 6917..6936 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7155..7259) /gene="bla" /label=AmpR promoter primer_bind complement(7334..7353) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagttaag cttggtacca tatggcggac gaggtggcgc tcgcccttca ggccgccggc 721 tccccttccg cggcggccgc catggaggcc gcgtcgcagc cggcggacga gccgctccgc 781 aagaggcccc gccgagacgg gcctggcctc gggcgcagcc cgggcgagcc gagcgcagca 841 gtggcgccgg cggccgcggg gtgtgaggcg gcgagcgccg cggccccggc ggcgctgtgg 901 cgggaggcgg caggggcggc ggcgagcgcg gagcgggagg ccccggcgac ggccgtggcc 961 ggggacggag acaatgggtc cggcctgcgg cgggagccga gggcggctga cgacttcgac 1021 gacgacgagg gcgaggagga ggacgaggcg gcggcggcag cggcggcggc agcgatcggc 1081 taccgaggtc catatacttt tgttcagcaa catctcatga ttggcaccga tcctcgaaca 1141 attcttaaag atttattacc agaaacaatt cctccacctg agctggatga tatgacgctg 1201 tggcagattg ttattaatat cctttcagaa ccaccaaagc ggaaaaaaag aaaagatatc 1261 aatacaattg aagatgctgt gaagttactg caggagtgta aaaagataat agttctgact 1321 ggagctgggg tttctgtctc ctgtgggatt cctgacttca gatcaagaga cggtatctat 1381 gctcgccttg cggtggactt cccagacctc ccagaccctc aagccatgtt tgatattgag 1441 tattttagaa aagacccaag accattcttc aagtttgcaa aggaaatata tcccggacag 1501 ttccagccgt ctctgtgtca caaattcata gctttgtcag ataaggaagg aaaactactt 1561 cgaaattata ctcaaaatat agataccttg gagcaggttg caggaatcca aaggatcctt 1621 cagtgtcatg gttcctttgc aacagcatct tgcctgattt gtaaatacaa agttgattgt 1681 gaagctgttc gtggagacat ttttaatcag gtagttcctc ggtgccctag gtgcccagct 1741 gatgagccac ttgccatcat gaagccagag attgtcttct ttggtgaaaa cttaccagaa 1801 cagtttcata gagccatgaa gtatgacaaa gatgaagttg acctcctcat tgttattgga 1861 tcttctctga aagtgagacc agtagcacta attccaagtt ctatacccca tgaagtgcct 1921 caaatattaa taaataggga acctttgcct catctacatt ttgatgtaga gctccttgga 1981 gactgcgatg ttataattaa tgagttgtgt cataggctag gtggtgaata tgccaaactt 2041 tgttgtaacc ctgtaaagct ttcagaaatt actgaaaaac ctccacgccc acaaaaggaa 2101 ttggttcatt tatcagagtt gccaccaaca cctcttcata tttcggaaga ctcaagttca 2161 cctgaaagaa ctgtaccaca agactcttct gtgattgcta cacttgtaga ccaagcaaca 2221 aacaacaatg ttaatgattt agaagtatct gaatcaagtt gtgtggaaga aaaaccacaa 2281 gaagtacaga ctagtaggaa tgttgagaac attaatgtgg aaaatccaga ttttaaggct 2341 gttggttcca gtactgcaga caaaaatgaa agaacttcag ttgcagaaac agtgagaaaa 2401 tgctggccta atagacttgc aaaggagcag attagtaagc ggcttgaggg taatcaatac 2461 ctgtttgtac caccaaatcg ttacatattc cacggtgctg aggtatactc agactctgaa 2521 gatgacgtct tgtcctctag ttcctgtggc agtaacagtg acagtggcac atgccagagt 2581 ccaagtttag aagaaccctt ggaagatgaa agtgaaattg aagaattcta caatggcttg 2641 gaagatgata cggagaggcc cgaatgtgct ggaggatctg gatttggagc tgatggaggg 2701 gatcaagagg ttgttaatga agctatagct acaagacagg aattgacaga tgtaaactat 2761 ccatcagaca aatcctcgag tctagggccc ttcgaacaaa aactcatctc agaagaggat 2821 ctgaatatgc ataccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc 2881 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2941 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 3001 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 3061 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 3121 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 3181 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 3241 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 3301 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 3361 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 3421 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 3481 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 3541 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3601 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3661 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3721 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3781 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3841 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3901 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 3961 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 4021 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 4081 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 4141 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 4201 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 4261 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 4321 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 4381 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 4441 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 4501 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 4561 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 4621 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 4681 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 4741 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 4801 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 4861 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4921 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 4981 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 5041 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 5101 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 5161 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 5221 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 5281 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 5341 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 5401 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 5461 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 5521 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 5581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 5641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 5701 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 5761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 5821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 5881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 5941 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 6001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 6061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 6121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 6181 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 6241 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 6301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 6361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 6421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 6481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 6541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 6601 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 6661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 6721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 6781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 6841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 6901 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 6961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 7021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 7081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 7141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 7201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 7261 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat 7321 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg 7381 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag 7441 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg 7501 atgtacgggc cagatatacg cgtt //