LOCUS Exported 6687 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression for structure determination; may not be full ORF. ACCESSION . VERSION . KEYWORDS RECQL5 (RECQL5A-c104) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6687) TITLE SGC Oxford 2017 REFERENCE 2 (bases 1 to 6687) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6687 /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 1552..1569 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1622..1640) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1636..1683 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1720..2175 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1807..1826) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2017..2038 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(2267..3082) /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 2990..3009 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3074..3093 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 3204..3792 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3693..3712 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3946..3963 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3978..4117 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4043..4062) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4203..4225 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4219..4410) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5219..6301) /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 6263..6282 /label=LacI-R /note="LacI, reverse primer" promoter complement(6302..6379) /gene="lacI" /label=lacI promoter primer_bind 6585..6604 /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 atccatggac cctgagcggc gagtccggag 181 tacgctgaag aaggtctttg ggtttgactc ttttaagacg cctttacagg agagtgcgac 241 catggctgta gtaaaaggta acaaggacgt ctttgtgtgc atgcccacag gggcaggaaa 301 atccctatgc tatcagctcc ctgctctgtt ggccaaaggc atcaccattg tagtctctcc 361 tctcattgct ttgattcagg accaagtgga ccacttgcta accctaaagg tacgagtaag 421 ttccctgaac tcgaagctct ctgcacagga aaggaaggag ctgcttgctg acctggagcg 481 agaaaagccc cagaccaaga ttctgtacat caccccagag atggcagctt catcctcctt 541 ccagcccacc ctgaactccc tggtgtcccg ccacctgctg tcttacttgg tggtggatga 601 agctcattgt gtttcccaat gggggcatga ctttcgtcct gactacttgc gtctgggtgc 661 cctgcgctcc cgcctgggac atgccccttg tgtggctctg accgccacag ccaccccaca 721 ggtccaagag gacgtgtttg ctgccctgca cctgaagaaa ccagttgcca tcttcaagac 781 tccctgcttc cgggccaacc tcttctatga tgtgcaattc aaggaactga tttctgatcc 841 ctatgggaac ctgaaggact tctgccttaa ggctcttgga caggaggctg ataaagggtt 901 atctggctgc ggcattgtgt actgcaggac tagagaggct tgtgaacagc tggccataga 961 gctcagctgc aggggtgtga acgccaaggc ttaccatgca gggctgaagg cctctgaaag 1021 aacgctggtg cagaacgact ggatggagga gaaggtccct gtaattgttg caaccattag 1081 ttttgggatg ggagtggata aagccaatgt caggtttgtc gcccattgga atattgccaa 1141 gtctatggct gggtactacc aggagtctgg ccgggctggc agggatggga agccttcctg 1201 gtgccgtctc tattactcca ggaatgaccg ggaccaagtc agcttcctga tcaggaagga 1261 agtagcaaaa ctccaggaaa agagaggaaa caaagcatct gataaagcca ctatcatggc 1321 ctttgatgcc ctggtgacct tctgtgaaga actggggtgc cgccatgccg ccattgccaa 1381 gtacttcggg gatgcgctgc ctgcctgcgc caaaggctgc gaccactgcc agaaccccac 1441 ggccgtgcgg aggcggctgg aggccttgga gcgcagcagc agctggtgac agtaaaggtg 1501 gatacggatc cgaattcgag ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac 1561 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc 1621 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt 1681 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat 1741 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 1801 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 1861 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 1921 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 1981 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 2041 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 2101 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 2161 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 2221 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat 2281 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa 2341 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat 2401 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct 2461 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga 2521 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt 2581 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac 2641 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca 2701 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct 2761 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga 2821 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct 2881 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat 2941 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc 3001 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg 3061 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt 3121 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 3181 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 3241 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 3301 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg 3361 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 3421 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 3481 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 3541 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 3601 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 3661 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 3721 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 3781 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 3841 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 3901 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc 3961 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca 4021 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata 4081 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc 4141 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt 4201 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct 4261 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc 4321 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat 4381 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt 4441 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga 4501 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg 4561 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt 4621 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc 4681 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc 4741 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc 4801 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac 4861 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg 4921 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc 4981 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca 5041 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc 5101 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg 5161 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc 5221 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 5281 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga 5341 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca 5401 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac 5461 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc 5521 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg 5581 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac 5641 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc 5701 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt 5761 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga 5821 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag 5881 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc 5941 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc 6001 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg 6061 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg 6121 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg 6181 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg 6241 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca 6301 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt 6361 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg 6421 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc 6481 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa 6541 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata 6601 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag 6661 aggatcgaga tctcgatccc gcgaaat //