LOCUS Exported 6385 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS KDM4D SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6385) TITLE SGC Oxford REFERENCE 2 (bases 1 to 6385) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6385 /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 1246..1263 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1316..1334) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1330..1377 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1414..1869 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1501..1520) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1711..1732 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1962..2777) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 2685..2704 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2769..2788 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2899..3487 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3388..3407 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3641..3658 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3673..3815 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3741..3760) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3901..3923 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3917..4108) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4917..5999) /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 5961..5980 /label=LacI-R /note="LacI, reverse primer" promoter complement(6000..6077) /gene="lacI" /label=lacI promoter primer_bind 6283..6302 /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 atccatggaa actatgaagt ctaaggccaa 181 ctgtgcccag aatccaaatt gtaacataat gatatttcat ccaaccaaag aggagtttaa 241 tgattttgat aaatatattg cttacatgga atcccaaggt gcacacagag ctggcttggc 301 taagataatt ccacccaaag aatggaaagc cagagagacc tatgataata tcagtgaaat 361 cttaatagcc actcccctcc agcaggtggc ctctgggcgg gcaggggtgt ttactcaata 421 ccataaaaaa aagaaagcca tgactgtggg ggagtatcgc catttggcaa acagtaaaaa 481 atatcagact ccaccacacc agaatttcga agatttggag cgaaaatact ggaagaaccg 541 catctataat tcaccgattt atggtgctga catcagtggc tccttgtttg atgaaaacac 601 taaacaatgg aatcttgggc acctgggaac aattcaggac ctgctggaaa aggaatgtgg 661 ggttgtcata gaaggcgtca atacacccta cttgtacttt ggcatgtgga aaaccacgtt 721 tgcttggcat acagaggaca tggaccttta cagcatcaac tacctgcacc ttggggagcc 781 caaaacttgg tatgtggtgc ccccagaaca tggccagcgc ctggaacgcc tggccaggga 841 gctcttccca ggcagttccc ggggttgtgg ggccttcctg cggcacaagg tggccctcat 901 ctcgcctaca gttctcaagg aaaatgggat tcccttcaat cgcataactc aggaggctgg 961 agagttcatg gtgacctttc cctatggcta ccatgctggc ttcaaccatg gtttcaactg 1021 cgcagaggcc atcaattttg ccactccgcg atggattgat tatggcaaaa tggcctccca 1081 gtgtagctgt ggggaggcaa gggtgacctt ttccatggat gccttcgtgc gcatcctgca 1141 acctgagcgc tatgacctgt ggaaacgtgg gcaagaccgg tgacagtaaa ggtggatacg 1201 gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca ccaccaccac 1261 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct 1321 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg 1381 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg 1441 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 1501 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 1561 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 1621 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 1681 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 1741 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt 1801 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt 1861 ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 1921 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc 1981 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc 2041 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg 2101 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca 2161 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc 2221 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca 2281 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat 2341 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac 2401 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat 2461 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa 2521 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct 2581 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc 2641 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta 2701 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc 2761 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt 2821 gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 2881 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 2941 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 3001 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc 3061 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 3121 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 3181 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 3241 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 3301 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 3361 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 3421 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 3481 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 3541 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 3601 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 3661 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg 3721 catatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca 3781 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg 3841 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct 3901 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc 3961 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt 4021 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt 4081 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca 4141 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg 4201 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg 4261 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc 4321 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg 4381 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc 4441 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat 4501 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga 4561 tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt 4621 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa 4681 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga 4741 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga 4801 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca 4861 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac 4921 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4981 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg 5041 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg 5101 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat 5161 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg 5221 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca 5281 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc 5341 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag 5401 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc 5461 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa 5521 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg 5581 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca 5641 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt 5701 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg 5761 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac 5821 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc 5881 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa 5941 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca 6001 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg 6061 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa 6121 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa 6181 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca 6241 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata 6301 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag 6361 gatcgagatc tcgatcccgc gaaat //