LOCUS pET-28a_(+)_IleRS 8083 bp ds-DNA circular 23-MAR-2021 DEFINITION . KEYWORDS "accession:addgene_vdb_2565" FEATURES Location/Qualifiers gene complement(717..980) /label="tet (300 - 563)" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 primer 909..928 /label="pBRrevBam_primer" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e promoter 1012..1030 /label="T7_promoter" /ApEinfo_revcolor=#f8d3a9 /ApEinfo_fwdcolor=#f8d3a9 regulatory 1030..1057 /label="lacO" /ApEinfo_revcolor=#b4abac /ApEinfo_fwdcolor=#b4abac feature 1075..1091 /label="T7_transl_en_RBS" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 misc_feature complement(1083..1107) /label="Tentative pET28 anneal FOR" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 misc_feature 1099..1101 /label="Start Codon" /ApEinfo_revcolor=#c7b0e3 /ApEinfo_fwdcolor=#c7b0e3 CDS 1099..3989 /label="Translation 1099-3989" misc_feature 1111..1128 /label="His Tag" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 tag 1138..1155 /label="Thrombin" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 primer 1158..1158 /label="Xpress_fwd_primer" /ApEinfo_revcolor=#ff9ccd /ApEinfo_fwdcolor=#ff9ccd misc_feature complement(1158..1158) /label="Tim Excises" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 gene 1159..3975 /label="ileS gene" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e CDS 1159..3975 /label="ileS CDS (isoleucyl-tRNA synthetase)" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 terminator 3983..4111 /label="T7_terminator" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 misc_feature 3987..3989 /label="Stop Codon" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 primer complement(4025..4043) /label="T7_Terminal_primer" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 gene complement(4671..5486) /label="KanR2" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 origin 5592..6211 /label="pBR322_origin" /ApEinfo_revcolor=#c6c9d1 /ApEinfo_fwdcolor=#c6c9d1 primer 6610..6632 /label="pGEX_3_primer" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 feature complement(6626..6817) /label="ROP" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 gene complement(7292..7567) /label="tet (576 - 851)" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 CDS join(7626..8083,1..634) /label="Translation 7626-634" regulatory complement(join(7626..8083,1..634)) /label="lacI" /ApEinfo_revcolor=#d59687 /ApEinfo_fwdcolor=#d59687 ORIGIN 1 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta 61 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca 121 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat 181 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat 241 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg 301 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt 361 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg 421 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg 481 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt 541 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga 601 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct 661 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct 721 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc 781 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg 841 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct 901 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg 961 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac 1021 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact 1081 ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg 1141 gtgccgcgcg gcagccatat gagtgactat aaatcaaccc tgaatttgcc ggaaacaggg 1201 ttcccgatgc gtggcgatct cgccaagcgc gaacccggaa tgctggcgcg ttggactgat 1261 gatgatctgt acggcatcat ccgtgcggct aaaaaaggca aaaaaacctt cattctgcat 1321 gatggccctc cttatgcgaa tggcagcatt catattggtc actcggttaa caagattctg 1381 aaagacatta tcgtgaagtc caaagggctt tccggttatg actcgccgta tgtgcctggc 1441 tgggactgcc acggtctgcc gatcgagctg aaagtcgagc aagaatacgg taagccgggt 1501 gagaaattca ccgccgccga gttccgcgcc aagtgccgcg aatacgcggc gacccaggtt 1561 gacggtcaac gcaaagactt tatccgtctg ggcgtgctgg gcgactggtc gcacccgtac 1621 ctgaccatgg acttcaaaac tgaagccaac atcatccgcg cgctgggcaa aatcatcggc 1681 aacggtcacc tgcacaaagg cgcgaagcca gttcactggt gcgttgactg ccgttctgcg 1741 ctggcggaag cggaagttga gtattacgac aaaacttctc cgtccatcga cgttgctttc 1801 caggcagtcg atcaggatgc actgaaagca aaatttgccg taagcaacgt taacggccca 1861 atctcgctgg taatctggac caccacgccg tggactctgc ctgccaaccg cgcaatctct 1921 attgcaccag atttcgacta tgcgctggtg cagatcgacg gtcaggccgt gattctggcg 1981 aaagatctgg ttgaaagcgt aatgcagcgt atcggcgtga ccgattacac cattctcggc 2041 acggtaaaag gtgcggagct tgagctgctg cgctttaccc atccgtttat gggcttcgac 2101 gttccggcaa tcctcggcga tcacgttacc ctggatgccg gtaccggtgc cgttcacacc 2161 gcgcctggcc acggcccgga cgactatgtg atcggtcaga aatacggcct ggaaaccgct 2221 aacccggttg gcccggacgg cacttatctg ccgggcactt atccgacgct ggatggcgtg 2281 aacgtcttca aagcgaacga catcgtcgtt gcgctgctgc aggaaaaagg cgcgctgctg 2341 cacgttgaga aaatgcagca cagctatccg tgctgctggc gtcacaaaac gccgatcatc 2401 ttccgcgcga cgccgcagtg gttcgtcagc atggatcaga aaggtctgcg tgcgcagtca 2461 ctgaaagaga tcaaaggcgt gcagtggatc ccggactggg gccaggcgcg tatcgagtcg 2521 atggttgcta accgtcctga ctggtgtatc tcccgtcagc gcacctgggg tgtaccgatg 2581 tcactgttcg tgcacaaaga cacggaagag ctgcatccgc gtacccttga actgatggaa 2641 gaagtggcaa aacgcgttga agtcgatggc atccaggcgt ggtgggatct cgatgcgaaa 2701 gagatcctcg gcgacgaagc tgatcagtac gtgaaagtgc cggacacatt ggatgtatgg 2761 tttgactccg gatctaccca ctcttctgtt gttgacgtgc gtccggaatt tgccggtcac 2821 gcagcggaca tgtatctgga aggttctgac caacaccgcg gctggttcat gtcttcccta 2881 atgatctcca ccgcgatgaa gggtaaagcg ccgtatcgtc aggtactgac ccacggcttt 2941 accgtggatg gtcagggccg caagatgtct aaatccatcg gcaataccgt ttcgccgcag 3001 gatgtgatga acaaactggg cgcggatatt ctgcgtctgt gggtggcatc aaccgactac 3061 accggtgaaa tggccgtttc tgacgagatc ctgaaacgtg ctgccgatag ctatcgtcgt 3121 atccgtaaca ccgcgcgctt cctgctggca aacctgaacg gttttgatcc agcaaaagat 3181 atggtgaaac cggaagagat ggtggtactg gatcgctggg ccgtaggttg tgcgaaagcg 3241 gcacaggaag acatcctcaa ggcgtacgaa gcatacgatt tccacgaagt ggtacagcgt 3301 ctgatgcgct tctgctccgt tgagatgggt tccttctacc tcgacatcat caaagaccgt 3361 cagtacaccg ccaaagcgga cagtgtggcg cgtcgtagct gccagactgc gctatatcac 3421 atcgcagaag cgctggtgcg ctggatggca ccaatcctct ccttcaccgc tgatgaagtg 3481 tggggctacc tgccgggcga acgtgaaaaa tacgtcttca ccggtgagtg gtacgaaggc 3541 ctgtttggcc tggcagacag tgaagcgatg aacgatgcgt tctgggacga gctgttgaaa 3601 gtgcgtggcg aagtgaacaa agtcattgag caagcgcgtg ccgacaagaa agtgggtggc 3661 tcgctggaag cggcagtaac cttgtatgca gaaccggaac tgtcggcgaa actgaccgcg 3721 ctgggcgatg aattacgatt tgtcctgttg acctccggcg ctaccgttgc agactataac 3781 gacgcacctg ctgatgctca gcagagcgaa gtactcaaag ggctgaaagt cgcgttgagt 3841 aaagccgaag gtgagaagtg cccacgctgc tggcactaca cccaggatgt cggcaaggtg 3901 gcggaacacg cagaaatctg cggccgctgt gtcagcaacg tcgccggtga cggtgaaaaa 3961 cgtaagtttg cctgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct 4021 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt 4081 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg 4141 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc 4201 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc 4261 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg 4321 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg 4381 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg 4441 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt 4501 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa 4561 tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 4621 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac 4681 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt 4741 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca 4801 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc 4861 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt 4921 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc 4981 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg 5041 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg 5101 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat 5161 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta 5221 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc 5281 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc 5341 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga 5401 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa 5461 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac 5521 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 5581 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 5641 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 5701 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 5761 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 5821 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 5881 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 5941 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 6001 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 6061 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 6121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 6181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 6241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 6301 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 6361 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 6421 ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 6481 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa 6541 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 6601 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 6661 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt 6721 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc 6781 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat 6841 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta 6901 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga 6961 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg 7021 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc 7081 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg 7141 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg 7201 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca 7261 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc 7321 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga 7381 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa 7441 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa 7501 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc 7561 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt 7621 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7681 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc 7741 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag 7801 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg 7861 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg 7921 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc 7981 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac 8041 atggcactcc agtcgccttc ccgttccgct atcggctgaa ttt //