LOCUS pET24_His-TEV-His 5294 bp DNA circular SYN 24-FEB-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5294) AUTHORS DF TITLE Direct Submission JOURNAL Exported Feb 19, 2024 from SnapGene Viewer 7.1.1 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5294 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 77..665 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 851..993 /label=bom /note="basis of mobility region from pBR322" CDS complement(1095..1286) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(2095..3177) /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" promoter complement(3178..3255) /gene="lacI" /label=lacI promoter promoter 3564..3582 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 3583..3607 /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 3622..3644 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" primer_bind complement(3637..3678) /label=pET24-HIS-TEV-r CDS 3661..3678 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind 3679..3723 /label=pET24-HIS-TEV-f CDS 3682..3702 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3718..3735 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 3802..3849 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 3886..4341 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(4434..5249) /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" ORIGIN 1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 841 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca 901 tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact 961 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac 1021 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc 1081 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg 1141 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc 1201 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta 1261 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg 1321 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa 1381 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac 1441 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca 1501 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac 1561 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag 1621 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc 1681 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc 1741 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg 1801 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc 1861 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc 1921 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg 1981 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc 2041 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg 2101 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 2161 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg 2221 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct 2281 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga 2341 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga 2401 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt 2461 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca 2521 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc 2581 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg 2641 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat 2701 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca 2761 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact 2821 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc 2881 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac 2941 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg 3001 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc 3061 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac 3121 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt 3181 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg 3241 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc 3301 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg 3361 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag 3421 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg 3481 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga 3541 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa 3601 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatggctagc 3661 catcatcacc atcatcacgt ggaaaacctg tactttcaag gtgcggccgc actcgagcac 3721 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct 3781 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg 3841 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg 3901 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 3961 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 4021 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 4081 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 4141 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 4201 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga 4261 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 4321 aaatattaac gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 4381 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa 4441 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat 4501 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg 4561 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat 4621 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc 4681 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta 4741 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga 4801 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac 4861 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct 4921 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga 4981 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg 5041 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct 5101 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg 5161 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag 5221 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca 5281 gacagtttta ttgt //