LOCUS Exported 5911 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION E.Coli produced SIN3A PAH1 purified by FPLC nickel column. ACCESSION . VERSION . KEYWORDS His-Sin3a-115-212(pL01) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5911) AUTHORS Chandru A, Bate N, Vuister GW, Cowley SM TITLE Sin3A recruits Tet1 to the PAH1 domain via a highly conserved Sin3-Interaction Domain. JOURNAL Sci Rep. 2018 Oct 2;8(1):14689. doi: 10.1038/s41598-018-32942-w. PUBMED 30279502 REFERENCE 2 (bases 1 to 5911) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5911 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 92..109 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 134..154 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 517..534 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 700..786 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 878..905 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind complement(959..977) /label=T7 Term /note="T7 terminator, reverse primer" terminator 973..1020 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1057..1512 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1144..1163) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1354..1375 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1627..2487) /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 2250..2269 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2488..2580) /gene="bla" /label=AmpR promoter rep_origin 2661..3249 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3150..3169 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3403..3420 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3435..3574 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3500..3519) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3660..3682 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3676..3867) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4439..5521) /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 5483..5502 /label=LacI-R /note="LacI, reverse primer" promoter complement(5522..5599) /gene="lacI" /label=lacI promoter primer_bind 5805..5824 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt 121 agatctgggt accgagaacc tgtacttcca atccatgcag cagcaatttc agaggctgaa 181 ggtggaggat gcgctatctt atcttgacca ggtgaagctg cagtttggta gtcagcctca 241 ggtctacaat gatttccttg acatcatgaa ggaatttaaa tctcagagca tcgacacccc 301 aggagtgatt agtcgtgtgt cccagctatt caaaggccac cccgatctga taatgggatt 361 caacaccttc ttgccccctg gctacaaaat tgaggtgcaa accaatgaca tggtgaatgt 421 gacaactcct ggccaggttc atcagattcc ctgacagtaa aggtggatac ggatccgaat 481 tcgagctccg tcgacaagct tgcggccgca ctcgagcacc accaccacca ccactaatgt 541 taattaagtt gggcgttcct aggctgataa aacagaattt gcctggcggc agtagcgcgg 601 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg 661 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag 721 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg 781 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca 841 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc 901 ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc 961 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg 1021 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa 1081 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 1141 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 1201 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 1261 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 1321 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 1381 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 1441 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 1501 cgtttacaat ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat 1561 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 1621 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 1681 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 1741 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 1801 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 1861 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 1921 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 1981 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 2041 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 2101 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 2161 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 2221 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 2281 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 2341 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 2401 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 2461 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt 2521 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 2581 taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 2641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 2701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 2761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 2821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 2881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 2941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 3001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 3061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 3121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 3181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 3241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 3301 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 3361 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 3421 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac 3481 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac 3541 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 3601 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 3661 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 3721 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc 3781 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 3841 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat 3901 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga 3961 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga 4021 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc 4081 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga 4141 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca 4201 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt 4261 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat 4321 catgctagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg 4381 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc 4441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 4501 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga 4561 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca 4621 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac 4681 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc 4741 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg 4801 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac 4861 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc 4921 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt 4981 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga 5041 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag 5101 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc 5161 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc 5221 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg 5281 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg 5341 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg 5401 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg 5461 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca 5521 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt 5581 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg 5641 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc 5701 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa 5761 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata 5821 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag 5881 aggatcgaga tcgatctcga tcccgcgaaa t //