LOCUS       pET28a-IleRS            8063 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Nucleus PURE: pET28a-IleRS in pET28a protein expression plasmid. See
            https://nucleus.bnext.bio/ for more details..
ACCESSION   pET28a-IleRS
VERSION     pET28a-IleRS
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..6498
                     /codon_start=1
                     /standard_name="IleRS"
                     /label="IleRS"
                     /translation="MSDYKSTLNLPETGFPMRGDLAKREPGMLARWTDDDLYGIIRAAK
                     KGKKTFILHDGPPYANGSIHIGHSVNKILKDIIVKSKGLSGYDSPYVPGWDCHGLPIEL
                     KVEQEYGKPGEKFTAAEFRAKCREYAATQVDGQRKDFIRLGVLGDWSHPYLTMDFKTEA
                     NIIRALGKIIGNGHLHKGAKPVHWCVDCRSALAEAEVEYYDKTSPSIDVAFQAVDQDAL
                     KAKFAVSNVNGPISLVIWTTTPWTLPANRAISIAPDFDYALVQIDGQAVILAKDLVESV
                     MQRIGVTDYTILGTVKGAELELLRFTHPFMGFDVPAILGDHVTLDAGTGAVHTAPGHGP
                     DDYVIGQKYGLETANPVGPDGTYLPGTYPTLDGVNVFKANDIVVALLQEKGALLHVEKM
                     QHSYPCCWRHKTPIIFRATPQWFVSMDQKGLRAQSLKEIKGVQWIPDWGQARIESMVAN
                     RPDWCISRQRTWGVPMSLFVHKDTEELHPRTLELMEEVAKRVEVDGIQAWWDLDAKEIL
                     GDEADQYVKVPDTLDVWFDSGSTHSSVVDVRPEFAGHAADMYLEGSDQHRGWFMSSLMI
                     STAMKGKAPYRQVLTHGFTVDGQGRKMSKSIGNTVSPQDVMNKLGADILRLWVASTDYT
                     GEMAVSDEILKRAADSYRRIRNTARFLLANLNGFDPAKDMVKPEEMVVLDRWAVGCAKA
                     AQEDILKAYEAYDFHEVVQRLMRFCSVEMGSFYLDIIKDRQYTAKADSVARRSCQTALY
                     HIAEALVRWMAPILSFTADEVWGYLPGEREKYVFTGEWYEGLFGLADSEAMNDAFWDEL
                     LKVRGEVNKVIEQARADKKVGGSLEAAVTLYAEPELSAKLTALGDELRFVLLTSGATVA
                     DYNDAPADAQQSEVLKGLKVALSKAEGEKCPRCWHYTQDVGKVAEHAEICGRCVSNVAG
                     DGEKRKFA"
     terminator      6572..6619
                     /label="T7 terminator"
                     /note="Color: #ffffff NCBI Feature Key: terminator
                     Transferred From: T7_terminator Transferred Similarity:
                     100.00%"
     rep_origin      6656..7111
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(7203..8018)
                     /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 gcaccatcac
     3661 caccatcatg gaggcggtgg atctatgtct gactacaagt caaccttaaa tctgccggaa
     3721 acgggtttcc cgatgcgggg cgacctggcg aaacgcgaac cgggcatgtt agcgcgttgg
     3781 acggacgatg atttgtatgg aatcatccgc gctgcgaaaa aaggtaagaa aacctttatt
     3841 cttcacgatg gccctccata tgcaaacggt tccatccaca ttgggcattc ggttaataaa
     3901 attctgaaag acatcatcgt gaaaagcaaa ggtttatccg gttacgatag cccgtatgtt
     3961 cctggatggg actgtcatgg tttgccgatc gagctgaagg tggagcagga atacggtaaa
     4021 cctggcgaaa aatttacagc agctgaattc cgcgcgaaat gccgtgaata cgcagccacg
     4081 caggtcgatg gccagcgcaa agactttatc cgcctggggg tactgggaga ctggtcacat
     4141 ccgtacctga ctatggactt caaaacagag gctaatatta ttcgtgcttt gggcaagatc
     4201 atcggtaatg gccacctcca taaaggggct aaaccggtgc attggtgtgt ggattgtcgc
     4261 tcggcgttgg cggaagcaga ggtagaatac tacgacaaaa cctccccgag cattgacgtg
     4321 gcttttcaag ccgttgacca ggatgctctg aaagcgaaat ttgcggtttc gaacgtgaac
     4381 ggtccaatct ctctggtcat ttggaccacc accccatgga ctctgcccgc caatcgtgcc
     4441 atctcgattg ctccagactt cgattacgct ctggtgcaga tcgacggtca agctgtgatc
     4501 ttggcgaaag accttgtgga gagtgttatg cagcgcattg gtgtgacaga ttacaccatt
     4561 ctgggtacgg tgaaaggtgc ggaactggaa ttacttcgtt ttacgcaccc ttttatgggg
     4621 tttgacgttc cagcgatctt aggcgatcat gttaccctgg acgcaggtac cggggccgtt
     4681 catacagccc caggtcacgg accggacgat tacgttattg ggcagaagta tggtctggaa
     4741 actgctaatc cggtcggccc cgatgggacg tacctgccag gcacctatcc gactttggac
     4801 ggcgttaatg tgtttaaagc gaacgacatc gttgttgcgc tgttacagga aaaaggtgcc
     4861 ctgctgcatg tggaaaagat gcaacactcg tatccgtgtt gctggcgcca caaaaccccg
     4921 atcatttttc gtgctactcc acagtggttt gtatccatgg accagaaagg gctgcgcgcc
     4981 caatcattaa aagagatcaa aggggttcag tggattcccg attggggcca ggcacgtatt
     5041 gagtctatgg ttgcaaatcg cccggattgg tgtatcagtc gccaacgcac atggggcgtt
     5101 ccgatgagcc tgtttgttca taaagacacc gaggagttgc atccgcgtac cttggagctg
     5161 atggaagaag tggcgaaacg cgtggaggtg gatggcatcc aggcgtggtg ggatttggat
     5221 gcaaaagaga ttctgggtga tgaagctgat caatacgtca aagtccccga cacgttagac
     5281 gtatggtttg attctggctc cactcacagc agcgttgttg acgttcggcc ggaatttgct
     5341 gggcacgcag ccgatatgta tttggaaggt tccgaccagc atcggggttg gtttatgtcg
     5401 tccctgatga ttagtaccgc catgaaagga aaagcacctt accgccaggt tttgactcat
     5461 gggtttacag tggatgggca ggggcgcaaa atgagtaaat ccattggcaa cacggtcagt
     5521 ccacaggatg tcatgaataa gctgggtgct gacattttgc gtctgtgggt ggcaagtacg
     5581 gactacaccg gtgaaatggc ggtttctgac gagattttga aacgtgccgc agattcctat
     5641 cgccgtattc gtaataccgc tcggttcctg ttagcgaatt tgaacggctt cgatcccgcg
     5701 aaagatatgg ttaagcccga ggaaatggtt gtgctggacc gctgggctgt agggtgcgca
     5761 aaggcagctc aggaggatat cctgaaagcg tacgaggcgt atgatttcca cgaggttgtg
     5821 cagcgtctga tgcgcttctg tagcgtggag atgggctcct tctatttgga tattattaaa
     5881 gatcgtcagt acacggcaaa agccgactcg gtggcccgcc gtagctgtca gaccgcgtta
     5941 taccacattg ccgaagcgct ggttcgttgg atggccccga ttttatcgtt cacagccgat
     6001 gaagtctggg gctatctgcc aggagagcgt gaaaaatacg tgtttacagg ggaatggtat
     6061 gaagggttgt ttggtctggc cgatagtgaa gccatgaacg atgccttctg ggatgaatta
     6121 ctgaaggtcc gtggcgaggt taataaggtg atcgaacagg ctcgcgcgga caagaaagtt
     6181 ggcggttccc tggaagctgc ggttacgtta tacgctgagc ccgaactgtc cgccaaactg
     6241 acggctttgg gggatgaact gcgttttgta ttgctgacat ctggggcaac ggtcgccgac
     6301 tataacgatg caccggccga tgctcagcag agcgaagtgc tgaaagggtt aaaagtagct
     6361 ttatcgaaag ctgaaggcga aaaatgtccg cgctgctggc actataccca agatgttggt
     6421 aaagtcgcag agcacgccga aatctgcggc cgctgtgtta gcaacgtggc cggcgacggc
     6481 gagaaacgta aatttgcctg aagagcttga tccggctgct aacaaagccc gaaaggaagc
     6541 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     6601 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     6661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6841 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6961 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     7021 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7081 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     7141 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     7201 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     7261 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     7321 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     7381 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     7441 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     7501 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     7561 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     7621 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     7681 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     7741 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     7801 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     7861 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     7921 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     7981 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     8041 atgtaagcag acagttttat tgt
//
