LOCUS       pET28a-MetRS            7277 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Nucleus PURE: pET28a-MetRS in pET28a protein expression plasmid. See
            https://nucleus.bnext.bio/ for more details..
ACCESSION   pET28a-MetRS
VERSION     pET28a-MetRS
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..5679
                     /codon_start=1
                     /standard_name="MetRS"
                     /label="MetRS"
                     /translation="MTQVAKKILVTCALPYANGSIHLGHMLEHIQADVWVRYQRMRGHE
                     VNFICADDAHGTPIMLKAQQLGITPEQMIGEMSQEHQTDFAGFNISYDNYHSTHSEENR
                     QLSELIYSRLKENGFIKNRTISQLYDPEKGMFLPDRFVKGTCPKCKSPDQYGDNCEVCG
                     ATYSPTELIEPKSVVSGATPVMRDSEHFFFDLPSFSEMLQAWTRSGALQEQVANKMQEW
                     FESGLQQWDISRDAPYFGFEIPNAPGKYFYVWLDAPIGYMGSFKNLCDKRGDSVSFDEY
                     WKKDSTAELYHFIGKDIVYFHSLFWPAMLEGSNFRKPSNLFVHGYVTVNGAKMSKSRGT
                     FIKASTWLNHFDADSLRYYYTAKLSSRIDDIDLNLEDFVQRVNADIVNKVVNLASRNAG
                     FINKRFDGVLASELADPQLYKTFTDAAEVIGEAWESREFGKAVREIMALADLANRYVDE
                     QAPWVVAKQEGRDADLQAICSMGINLFRVLMTYLKPVLPKLTERAEAFLNTELTWDGIQ
                     QPLLGHKVNPFKALYNRIDMRQVEALVEASKEEVKAAAAPVTGPLADDPIQETITFDDF
                     AKVDLRVALIENAEFVEGSDKLLRLTLDLGGEKRNVFSGIRSAYPDPQALIGRHTIMVA
                     NLAPRKMRFGISEGMVMAAGPGGKDIFLLSPDAGAKPGHQVK"
     CDS             5680..5694
                     /codon_start=1
                     /label="Linker"
                     /translation="GGGGS"
     CDS             5695..5712
                     /codon_start=1
                     /label="His6"
                     /translation="HHHHHH"
     terminator      5786..5833
                     /label="T7 terminator"
                     /note="Color: #ffffff NCBI Feature Key: terminator
                     Transferred From: T7_terminator Transferred Similarity:
                     100.00%"
     rep_origin      5870..6325
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(6417..7232)
                     /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 gacccaagtt
     3661 gcgaaaaaaa tcctggtcac gtgcgccctg ccgtacgcga acggatctat ccatttgggc
     3721 cacatgcttg aacatattca ggcggatgtt tgggtgcgct accagcgtat gcgtgggcat
     3781 gaagtgaact tcatttgcgc ggacgatgcg catggtacac ccattatgct gaaggcgcag
     3841 caattaggga ttacccccga acagatgatc ggagaaatgt ctcaagagca tcaaactgat
     3901 tttgcgggct tcaacattag ttacgataac tatcatagca ctcattctga ggaaaatcgt
     3961 cagctgagtg agctgatcta ttcacgcctg aaggaaaacg gtttcatcaa aaaccgcacg
     4021 atctcccagc tgtacgatcc agaaaaaggt atgtttttac cggaccgttt cgttaaaggt
     4081 acctgtccga aatgcaaatc gccggatcaa tatggtgata actgtgaagt ctgtggtgct
     4141 acttatagcc cgacagagct gattgagcca aaaagcgtag tctcaggtgc cactcctgtg
     4201 atgcgtgact ctgagcactt tttcttcgac ctgccgagct tcagtgaaat gttacaggcc
     4261 tggacccgtt cgggcgcgct gcaggaacag gtggcgaata aaatgcaaga atggtttgaa
     4321 tccggcctgc agcagtggga tatttcgcgc gacgctccct atttcggctt tgagattccg
     4381 aacgcgcccg gtaagtattt ctatgtatgg cttgatgccc cgattggcta tatggggagc
     4441 ttcaaaaact tatgcgataa acgtggcgat agtgttagtt ttgatgaata ctggaaaaag
     4501 gatagcaccg cggaacttta tcatttcatt ggcaaagaca ttgtttattt ccacagtctg
     4561 ttctggccag caatgttaga aggcagcaac tttcgcaaac caagcaacct gttcgttcac
     4621 ggctatgtta ccgtgaatgg tgcaaaaatg tccaagtcac gcggaacatt tattaaagct
     4681 agtacgtggc ttaaccactt cgatgcggat tctctgcggt actactacac agctaagctt
     4741 tctagccgca tcgatgatat cgatcttaat ttagaagatt ttgtccagcg cgtgaacgca
     4801 gatattgtga ataaggtcgt taacttagcc agtcgtaatg ccgggttcat caataaacgg
     4861 tttgacggtg tcctggctag cgagcttgca gatccgcaac tttataagac ttttaccgat
     4921 gcggcggaag ttattggtga ggcatgggaa tcacgtgaat tcggaaaggc cgtccgtgaa
     4981 attatggcgc tggcagacct ggcgaaccgc tacgtggatg aacaagcgcc gtgggttgtt
     5041 gccaagcagg aaggccgcga cgctgacttg caggctattt gctctatggg cattaattta
     5101 ttccgtgttc tgatgacata tctgaaaccc gtgctgccta agttgacgga gcgcgcggag
     5161 gcctttctga ataccgaact gacgtgggat ggcattcagc agccgctgct gggtcataaa
     5221 gtgaacccgt tcaaagcgtt gtacaaccgc attgatatgc gccaagtgga agcgttggtt
     5281 gaagcctcaa aagaggaagt gaaagcggcg gccgctcccg ttactggccc gctggcggat
     5341 gatccaatcc aagagacaat cacctttgat gattttgcca aagttgattt acgcgtagcg
     5401 ttaattgaga acgcggaatt cgtggagggg tcggataaac tgctgcgtct gacgctggat
     5461 ctggggggcg agaaacgcaa tgtattctca gggattcgct ccgcgtatcc ggatccacag
     5521 gcgcttatcg gacgtcacac gatcatggtg gctaatttgg cccctcgcaa aatgcgcttc
     5581 ggtatttctg aaggcatggt catggccgcg ggacccggcg gcaaggacat tttcctgtta
     5641 tcgcctgacg cgggcgcgaa accgggacac caagtaaagg gaggcggtgg atctcaccat
     5701 caccaccatc attgaagagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     5761 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     5821 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     5881 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5941 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6001 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6061 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6121 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6181 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6241 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6301 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6361 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag
     6421 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     6481 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     6541 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     6601 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     6661 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     6721 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     6781 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     6841 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     6901 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     6961 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     7021 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     7081 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     7141 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     7201 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     7261 gcagacagtt ttattgt
//
