LOCUS       pET28a-GlyRS            8276 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Nucleus PURE: pET28a-GlyRS in pET28a protein expression plasmid. See
            https://nucleus.bnext.bio/ for more details..
ACCESSION   pET28a-GlyRS
VERSION     pET28a-GlyRS
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      77..665
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        77..105
                     /label="RNAII promoter"
                     /note="Color: #ffffff NCBI Feature Key: promoter NCBI Join
                     Type: join Transferred From: RNAII_promoter Transferred
                     Similarity: 100.00%"
     misc_RNA        113..665
                     /label="RNAII transcript (complementary strand)"
                     /note="RNAII transcript (complementary strand)"
     misc_RNA        complement(116..223)
                     /label="RNAI transcript"
                     /note="RNAI transcript"
     promoter        complement(230..259)
                     /label="RNA I promoter"
                     /note="Color: #ffffff NCBI Feature Key: promoter NCBI Join
                     Type: join Transferred From: RNA_I_promoter Transferred
                     Similarity: 100.00%"
     terminator      669..712
                     /label="rop/bom terminator"
     misc_binding    752..760
                     /label="dnaA binding site"
                     /bound_moiety="dnaA"
     misc_feature    785..848
                     /label="L-strand Y effector site"
                     /note="L-strand Y effector site"
     misc_feature    851..990
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     misc_feature    1031..1187
                     /label="H-strand Y effector site"
                     /note="H-strand Y effector site"
     CDS             complement(1092..1283)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     RBS             1284..1293
     misc_feature    complement(1359..1377)
                     /label="ROP calculated promoter"
     CDS             complement(2092..3174)
                     /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(3175..3252)
                     /label="lacI promoter"
                     /note="Color: #ffffff NCBI Feature Key: promoter
                     Transferred From: lacI_promoter Transferred Similarity:
                     100.00%"
     promoter        3561..3579
                     /label="T7_promoter"
     protein_bind    3580..3604
                     /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             3613..3645
                     /label="UTR1"
                     /note="note: UTR1 NCBI Feature Key: misc_feature Color:
                     #cc0000 modified by: Andrey Transferred From: U4m - UTR1
                     Transferred Similarity: 100.00%"
     CDS             3649..3651
                     /codon_start=1
                     /label="Start"
                     /translation="M"
     CDS             3652..3669
                     /codon_start=1
                     /label="His6"
                     /translation="HHHHHH"
     CDS             3670..3684
                     /codon_start=1
                     /label="Linker"
                     /translation="GGGGS"
     CDS             3685..4596
                     /codon_start=1
                     /standard_name="GlyQ"
                     /label="GlyQ"
                     /translation="MQKFDTRTFQGLILTLQDYWARQGCTIVQPLDMEVGAGTSHPMTC
                     LRELGPEPMAAAYVQPSRRPTDGRYGENPNRLQHYYQFQVVIKPSPDNIQELYLGSLKE
                     LGMDPTIHDIRFVEDNWENPTLGAWGLGWEVWLNGMEVTQFTYFQQVGGLECKPVTGEI
                     TYGLERLAMYIQGVDSVYDLVWSDGPLGKTTYGDVFHQNEVEQSTYNFEYADVDFLFTC
                     FEQYEKEAQQLLALENPLPLPAYERILKAAHSFNLLDARKAISVTERQRYILRIRTLTK
                     AVAEAYYASREALGFPMCNKDK"
     CDS             4612..6678
                     /codon_start=1
                     /standard_name="GlyS"
                     /label="GlyS"
                     /translation="MSEKTFLVEIGTEELPPKALRSLAESFAANFTAELDNAGLAHGTV
                     QWFAAPRRLALKVANLAEAQPDREIEKRGPAIAQAFDAEGKPSKAAEGWARGCGITVDQ
                     AERLTTDKGEWLLYRAHVKGESTEALLPNMVATSLAKLPIPKLMRWGASDVHFVRPVHT
                     VTLLLGDKVIPATILGIQSDRVIRGHRFMGEPEFTIDNADQYPEILRERGKVIADYEER
                     KAKIKADAEEAARKIGGNADLSESLLEEVASLVEWPVVLTAKFEEKFLAVPAEALVYTM
                     KGDQKYFPVYANDGKLLPNFIFVANIESKDPQQIISGNEKVVRPRLADAEFFFNTDRKK
                     RLEDNLPRLQTVLFQQQLGTLRDKTDRIQALAGWIAEQIGADVNHATRAGLLSKCDLMT
                     NMVFEFTDTQGVMGMHYARHDGEAEDVAVALNEQYQPRFAGDDLPSNPVACALAIADKM
                     DTLAGIFGIGQHPKGDKDPFALRRAALGVLRIIVEKNLNLDLQTLTEEAVRLYGDKLTN
                     ANVVDDVIDFMLGRFRAWYQDEGYTVDTIQAVLARRPTRPADFDARMKAVSHFRTLDAA
                     AALAAANKRVSNILAKSDEVLSDRVNASTLKEPEEIKLAMQVVVLRDKLEPYFTEGRYQ
                     DALVELAELREPVDAFFDKVMVMVDDKELRINRLTMLEKLRELFLRVADISLLQ"
     CDS             6679..6693
                     /codon_start=1
                     /label="Linker"
                     /translation="GGGGS"
     CDS             6694..6711
                     /codon_start=1
                     /label="His6"
                     /translation="HHHHHH"
     terminator      6785..6832
                     /label="T7 terminator"
                     /note="Color: #ffffff NCBI Feature Key: terminator
                     Transferred From: T7_terminator Transferred Similarity:
                     100.00%"
     rep_origin      6869..7324
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(7416..8231)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label="KanR"
                     /note="confers resistance to kanamycin"
                     /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 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
      961 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     1021 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     1081 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     1141 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     1201 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     1261 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     1321 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     1381 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     1441 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     1501 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     1561 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     1621 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     1681 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     1741 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     1801 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     1861 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     1921 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     1981 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     2041 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     2101 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2161 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     2221 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     2281 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     2341 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     2401 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     2461 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     2521 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     2581 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     2641 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     2701 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     2761 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     2821 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     2881 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     2941 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     3001 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     3061 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     3121 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     3181 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     3241 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     3301 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     3361 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     3421 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     3481 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     3541 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     3601 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccaat gcatcaccat
     3661 catcaccacg gggggggcgg cagcatgcag aagtttgaca ctcgtacctt tcaaggactg
     3721 attctgactt tacaggatta ctgggctcgc cagggttgta ccatcgtcca gccgctggat
     3781 atggaagtcg gggcgggtac atcacatccg atgacctgct tgcgtgaact gggcccagag
     3841 ccgatggccg cagcgtacgt acagccgtca cgtcgtccca ccgacggtcg ctatggcgag
     3901 aacccaaatc gcctgcaaca ttattatcag tttcaggttg tcattaaacc gagccctgat
     3961 aacattcagg aattgtacct gggatcttta aaagaattag gcatggaccc gaccatccat
     4021 gatattcgct ttgtggaaga taactgggaa aacccaacgc tgggcgcgtg gggactgggt
     4081 tgggaagtgt ggctgaacgg catggaggtt actcaattta cctactttca acaggtcggt
     4141 ggcctggagt gcaaaccggt gaccggtgaa attacctatg gtctggaacg tcttgctatg
     4201 tatattcagg gcgtggattc cgtatacgat ctggtttggt ctgatggtcc gctgggtaaa
     4261 accacctatg gagatgtttt ccaccagaac gaagtagagc aaagtaccta taactttgaa
     4321 tacgcagatg tagacttttt atttacttgc ttcgaacaat acgagaagga agcacagcag
     4381 ttgctggccc tggaaaaccc gctgccgctg ccggcctatg aacgtatcct gaaggcggcg
     4441 cattcgttta atctgctgga cgcccgcaaa gcaatttcag tgacggaacg ccagcgttat
     4501 attctgcgta tccggaccct gacaaaggct gtggctgaag catattatgc atcacgtgaa
     4561 gcattgggct ttccgatgtg taataaagat aagtgaatta aagaggagaa aatgagtgaa
     4621 aaaacgttct tagttgaaat cggaaccgaa gagttgcctc cgaaggcgtt acgtagtctg
     4681 gcggaaagct ttgctgcgaa ctttaccgcg gagttggata acgccggcct ggcacatgga
     4741 actgtgcaat ggttcgcggc accgcgtcgc ttagcactta aggtggccaa tttggcagaa
     4801 gcccagcctg atcgcgaaat tgaaaaacgt ggtcccgcca ttgctcaggc attcgacgca
     4861 gaagggaagc catctaaagc ggctgaaggt tgggctcgtg gatgtggcat tactgtcgac
     4921 caagcggaac gcttaacgac tgataaaggt gaatggctgt tataccgcgc gcatgttaag
     4981 ggcgaaagca ccgaagcgct gttacctaac atggtcgcca cgtcactggc caaactgccg
     5041 attcctaaat taatgcgttg gggcgcaagt gatgtgcact ttgtacgtcc ggtgcacact
     5101 gtcaccctgt tgctgggtga taaagtgatc cctgccacca ttcttggaat ccagagtgac
     5161 cgcgtgattc gtggtcaccg ttttatgggt gaaccggaat tcacgatcga caatgctgac
     5221 cagtacccgg aaatcctgcg tgagcgcggt aaagttatcg ccgactacga agaacgtaaa
     5281 gctaaaatta aagccgacgc cgaagaagcc gcgcgcaaaa ttggtggcaa tgcggactta
     5341 agtgagagtt tgctggagga agtggcctca ttagtggagt ggcctgtggt tctgacagcg
     5401 aaatttgaag aaaagtttct tgctgtgcct gccgaggctt tagtttacac catgaagggc
     5461 gaccagaaat attttccggt gtacgccaac gacgggaagt tactgccaaa cttcattttt
     5521 gtcgctaata ttgagtcgaa agatccgcag cagattattt ctggtaacga aaaagtagtt
     5581 cgtccgcgcc tggccgacgc tgaatttttc tttaatactg atcgtaaaaa acgcttagaa
     5641 gataacctgc cccgtctgca gaccgtcctg tttcaacagc agcttggtac tttacgggat
     5701 aaaacagatc gcattcaggc gctggctggc tggatcgcgg agcagatcgg cgccgatgtg
     5761 aaccacgcta cacgtgccgg tctgcttagc aagtgcgatc tgatgactaa catggtattc
     5821 gagttcactg atacgcaggg cgtcatgggc atgcactatg cgcgtcatga tggcgaggcg
     5881 gaggacgtcg ctgtcgcatt gaacgaacag taccagcctc gtttcgccgg agatgatctg
     5941 ccgtcgaatc cggttgcttg tgcactggcg atcgcggata aaatggatac attagcgggg
     6001 atctttggca ttggtcagca ccccaaaggc gacaaagacc cgttcgctct gcgtcgtgcc
     6061 gcgctgggcg ttctgcgtat tattgtagaa aaaaatttga atctggactt acagaccctg
     6121 acagaagagg ctgtgcgcct gtatggggac aaattaacca atgcgaacgt ggttgatgac
     6181 gttattgact ttatgttagg acggtttcgc gcgtggtatc aggatgaagg ctatacggtg
     6241 gatacgattc aagccgtctt agcacgccgt cctacccgcc ccgcagactt cgacgcgcgc
     6301 atgaaagcgg tcagtcattt tcgcaccttg gatgcagccg ccgcccttgc cgccgcgaac
     6361 aaacgtgtgt ccaatattct ggcgaaatca gatgaagtgt tatcagatcg ggtgaatgca
     6421 tcaaccctta aagaacccga agaaatcaag cttgcaatgc aagttgttgt tcttcgcgac
     6481 aagctggagc catactttac tgagggtcgc tatcaggatg ctctggttga actggcggag
     6541 ctgcgcgaac cggtggacgc tttctttgac aaggttatgg tgatggtcga tgataaggaa
     6601 ctgcgcatca accgccttac aatgctggag aaattgcgtg aactgtttct gcgcgtcgca
     6661 gatatcagtt tgcttcaggg aggcggtgga tctcaccatc accaccatca ttgaagagct
     6721 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     6781 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     6841 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     6901 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     6961 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     7021 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     7081 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     7141 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     7201 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     7261 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     7321 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     7381 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     7441 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     7501 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     7561 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     7621 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     7681 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     7741 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     7801 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     7861 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     7921 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     7981 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     8041 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     8101 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     8161 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     8221 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgt
//
