LOCUS pET-28a_(+)_GlyRS 8257 bp ds-DNA circular 23-MAR-2021 DEFINITION . KEYWORDS "creator:Qiagen" "marker:KanR, CmR" "organism:Escherichia coli" 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..4163 /label="Translation 1099-4163" misc_feature 1111..1128 /label="His Tag" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 misc_feature 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 primer 1159..1185 /label="GlyN FOR" /note="sequence: agcagcggcctggtgccgcgcggcagccatatgcaaaagtttgataccaggaccttc" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 CDS 1159..2070 /label="glyQ CDS (glycine tRNA synthetase, alpha subunit)" /ApEinfo_revcolor=#c6c9d1 /ApEinfo_fwdcolor=#c6c9d1 gene 1159..2070 /label="glyQ gene" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 CDS 2080..4149 /label="glyS CDS (glycine tRNA synthetase, beta subunit)" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e gene 2080..4149 /label="glyS gene" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e primer complement(4121..4142) /label="GlyN REV" /note="sequence: cttcctttcgggctttgttagcagccggatcttattgcaacagcgaaatatccgcaacgc" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 terminator 4157..4285 /label="T7_terminator" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 misc_feature 4161..4163 /label="Stop Codon" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 primer complement(4199..4217) /label="T7_Terminal_primer" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 gene complement(4845..5660) /label="KanR2" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 origin 5766..6385 /label="pBR322_origin" /ApEinfo_revcolor=#c6c9d1 /ApEinfo_fwdcolor=#c6c9d1 primer 6784..6806 /label="pGEX_3_primer" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 feature complement(6800..6991) /label="ROP" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 gene complement(7466..7741) /label="tet (576 - 851)" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 regulatory complement(join(7800..8257,1..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 gcaaaagttt gataccagga ccttccaggg cttgatcctg 1201 accttacagg attactgggc tcgccagggc tgcaccattg ttcaaccatt ggacatggaa 1261 gtcggcgcgg gaacctctca cccaatgacc tgtctgcgcg agctggggcc agaaccgatg 1321 gcggctgctt atgttcagcc ttctcgtcgc ccgaccgatg gtcgctacgg cgaaaacccc 1381 aaccgtttac agcactacta tcagttccag gtggtcatta agccatcgcc ggacaatatt 1441 caggagctgt acctcggttc tctgaaagag ctgggcatgg acccgactat tcacgacatc 1501 cgtttcgtgg aagataactg ggaaaacccg acgctgggtg cctggggact gggctgggaa 1561 gtgtggctga acggcatgga agtgacgcag ttcacttact tccagcaggt tggtggtctg 1621 gagtgtaaac cggttaccgg cgagatcacc tacggtctgg aacgtctggc catgtacatt 1681 cagggcgtag acagcgttta cgacctggtc tggagcgacg gcccgctggg taaaaccacc 1741 tacggcgacg tgttccatca gaacgaagtg gagcagtcca cttacaactt cgaatacgcg 1801 gatgtggact tcctgttcac ctgcttcgag cagtacgaga aagaagcgca gcagctgctg 1861 gcgctggaaa atccgctgcc gctgccagcc tacgagcgta ttctgaaagc cgcccacagc 1921 ttcaacctgc tggatgcgcg taaagccatc tccgtcaccg agcgtcagcg ctacattctg 1981 cgcattcgca ccctgaccaa agcagtggca gaagcatact acgcttcccg tgaagccctc 2041 ggcttcccga tgtgcaacaa agataagtaa gaggcggcta tgtctgagaa aacttttctg 2101 gtggaaatcg gcactgaaga gctgccacca aaagcactgc gcagcctggc tgagtccttt 2161 gctgcgaact ttactgcgga gctggataac gctggcctcg cacacggcac cgttcaatgg 2221 tttgctgctc cgcgtcgtct ggcgctgaaa gtagctaacc tggcggaagc gcaaccggat 2281 cgtgaaatcg aaaaacgcgg cccggcgatt gcccaggcgt tcgacgctga aggcaaaccg 2341 agcaaagcgg cagaaggttg ggcgcgtggt tgcggtatta ccgttgacca ggctgagcgt 2401 ctgactaccg ataaaggcga atggctgctg tatcgcgccc atgtgaaggg cgaaagcacc 2461 gaagcactgc tgccgaatat ggttgcgact tctctggcga aactgccgat cccgaaactg 2521 atgcgttggg gcgcaagcga cgtgcacttc gtgcgtccgg tgcacaccgt gaccctgctg 2581 ctgggcgaca aagtcattcc ggcaaccatt ctgggcattc agtccgatcg cgtgattcgc 2641 ggccaccgct ttatgggcga gccggaattc accatcgata acgccgatca gtatccggaa 2701 attctgcgtg agcgtgggaa agtcatcgcc gattacgaag aacgtaaggc gaagattaaa 2761 gccgatgccg aagaagcagc gcgtaagatt ggcggtaacg ctgacttaag cgaaagcctg 2821 ctggaagaag tggcttcgct ggtggagtgg ccggtcgttc tgaccgcaaa attcgaagag 2881 aaattcctcg cggtgccggc tgaagcgctg gtttacacca tgaaaggtga ccagaaatac 2941 ttcccggtgt atgcgaacga cggcaaactg ctgccgaact ttatcttcgt tgccaacatc 3001 gaatcgaaag atccgcagca gattatctcc ggtaacgaga aagtcgttcg tccgcgtctg 3061 gcggatgccg agttcttctt caacaccgac cgtaaaaaac gtcttgaaga taacctgccg 3121 cgcctgcaaa ccgtgttgtt ccagcaacag ttggggacgc tgcgcgacaa aactgaccgc 3181 atccaggcgc tggctggctg gattgctgaa cagattggcg ctgacgttaa ccacgctacc 3241 cgtgcgggtc tgctgtctaa gtgcgacctg atgaccaaca tggtcttcga gttcaccgac 3301 acccagggcg ttatggggat gcactatgcg cgtcacgatg gcgaagcgga agatgtcgcg 3361 gtggcgctga atgagcagta tcagccgcgt tttgctggtg atgacctgcc gtccaaccca 3421 gtagcttgtg cgctggcgat tgctgacaag atggataccc tggcgggtat cttcggtatc 3481 ggtcagcatc cgaaaggcga caaagacccg tttgcgctgc gtcgtgccgc gcttggcgtg 3541 ctgcgaatta tcgttgagaa gaacctcaac cttgatctgc aaacgctgac cgaagaagcg 3601 gtgcgtctgt atggcgataa gctgactaat gccaacgtag ttgatgatgt tatcgacttt 3661 atgctcggtc gcttccgcgc ctggtatcag gacgaaggtt ataccgttga caccatccag 3721 gcggtactgg cgcgtcgtcc gactcgtccg gctgatttcg atgcccgtat gaaagcggta 3781 tcgcatttcc gtaccctgga tgcagctgct gcactggcgg cggcgaacaa acgtgtatct 3841 aacattctgg cgaaatctga cgaagtgctg agcgaccgcg tgaatgcctc taccctgaaa 3901 gagccggaag aaattaaact ggcgatgcag gttgtggtgc tacgtgacaa gctggagccg 3961 tactttacgg aaggtcgtta ccaggatgcg ctggtcgaac tggctgagct gcgtgaaccg 4021 gttgatgctt tcttcgataa agtgatggtc atggttgatg acaaagaatt gcgtatcaac 4081 cgtctgacca tgctggagaa actgcgcgaa ctgttcctgc gcgttgcgga tatttcgctg 4141 ttgcaataag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc 4201 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg 4261 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa 4321 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 4381 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 4441 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 4501 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 4561 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 4621 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 4681 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 4741 cgtttacaat ttcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 4801 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg 4861 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa 4921 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc 4981 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg 5041 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat 5101 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca 5161 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga 5221 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg 5281 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg 5341 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata 5401 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca 5461 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg 5521 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat 5581 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt 5641 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt 5701 attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 5761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 5821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 5881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta 5941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 6001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 6061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 6121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 6181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 6241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 6301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 6361 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 6421 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 6481 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 6541 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca 6601 ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat 6661 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg 6721 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg 6781 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc 6841 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg 6901 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca 6961 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt 7021 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg 7081 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc 7141 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg 7201 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg 7261 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg 7321 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt 7381 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca 7441 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac 7501 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg 7561 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc 7621 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg 7681 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg 7741 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct 7801 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7861 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag 7921 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc 7981 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa 8041 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc 8101 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc 8161 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca 8221 ctccagtcgc cttcccgttc cgctatcggc tgaattt //