LOCUS       pET28a-ArgRS            6996 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Nucleus PURE: pET28a-ArgRS in pET28a protein expression plasmid. See
            https://nucleus.bnext.bio/ for more details..
ACCESSION   pET28a-ArgRS
VERSION     pET28a-ArgRS
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             3648..3650
                     /codon_start=1
                     /label="Start"
                     /translation="M"
     CDS             3651..3659
                     /codon_start=1
                     /label="Linker"
                     /translation="GSS"
     CDS             3660..3677
                     /codon_start=1
                     /label="His6"
                     /translation="HHHHHH"
     CDS             3678..3686
                     /codon_start=1
                     /label="Linker"
                     /translation="SSG"
     CDS             3687..3704
                     /codon_start=1
                     /label="Thrombin"
                     /translation="LVPRGS"
     gene            3708..5441
                     /label="argS gene"
     CDS             3708..5438
                     /codon_start=1
                     /label="ArgRS"
                     /translation="MNIQALLSEKVRQAMIAAGAPADCEPQVRQSAKVQFGDYQANGMM
                     AVAKKLGMAPRQLAEQVLTHLDLNGIASKVEIAGPGFINIFLDPAFLAEHVQQALASDR
                     LGVATPEKQTIVVDYSAPNVAKEMHVGHLRSTIIGDAAVRTLEFLGHKVIRANHVGDWG
                     TQFGMLIAWLEKQQQENAGEMELADLEGFYRDAKKHYDEDEEFAERARNYVVKLQSGDE
                     YFREMWRKLVDITMTQNQITYDRLNVTLTRDDVMGESLYNPMLPGIVADLKAKGLAVES
                     EGATVVFLDEFKNKEGEPMGVIIQKKDGGYLYTTTDIACAKYRYETLHADRVLYYIDSR
                     QHQHLMQAWAIVRKAGYVPESVPLEHHMFGMMLGKDGKPFKTRAGGTVKLADLLDEALE
                     RARRLVAEKNPDMPADELEKLANAVGIGAVKYADLSKNRTTDYIFDWDNMLAFEGNTAP
                     YMQYAYTRVLSVFRKAEIDEEQLAAAPVIIREDREAQLAARLLQFEETLTVVAREGTPH
                     VMCAYLYDLAGLFSGFYEHCPILSAENEEVRNSRLKLAQLTAKTLKLGLDTLGIETVER
                     M"
     terminator      5505..5552
                     /label="T7 terminator"
                     /note="Color: #ffffff NCBI Feature Key: terminator
                     Transferred From: T7_terminator Transferred Similarity:
                     100.00%"
     rep_origin      5589..6044
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(6136..6951)
                     /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 atataccatg ggcagcagcc
     3661 atcatcatca tcatcacagc agcggcctgg tgccgcgcgg cagccatatg aatattcagg
     3721 ctcttctctc agaaaaagtc cgtcaggcca tgattgcggc aggcgcgcct gcggattgcg
     3781 aaccgcaggt tcgtcagtca gcaaaagttc agttcggcga ctatcaggct aacggcatga
     3841 tggcagttgc taaaaaactg ggtatggcac cgcgacaatt agcagagcag gtgctgactc
     3901 atctggatct taacggtatc gccagcaaag ttgagatcgc cggtccaggc tttatcaaca
     3961 ttttccttga tccggcattc ctggctgaac atgttcagca ggcgctggcg tccgatcgtc
     4021 tcggtgttgc tacgccagaa aaacagacca ttgtggttga ctactctgcg ccaaacgtgg
     4081 cgaaagagat gcatgtcggt cacctgcgct ctaccattat tggtgacgca gcagtgcgta
     4141 ctctggagtt cctcggtcac aaagtgattc gcgcaaacca cgtcggcgac tggggcactc
     4201 agttcggtat gctgattgca tggctggaaa agcagcagca ggaaaacgcc ggtgaaatgg
     4261 agctggctga ccttgaaggt ttctaccgcg atgcgaaaaa gcattacgat gaagatgaag
     4321 agttcgccga gcgcgcacgt aactacgtgg taaaactgca aagcggtgac gaatatttcc
     4381 gcgagatgtg gcgcaaactg gtcgacatca ccatgacgca gaaccagatc acctacgatc
     4441 gtctcaacgt gacgctgacc cgtgatgacg tgatgggcga aagcctctac aacccgatgc
     4501 tgccaggaat tgtggcggat ctcaaagcca aaggtctggc agtagaaagc gaaggggcga
     4561 ccgtcgtatt ccttgatgag tttaaaaaca aggaaggcga accgatgggc gtgatcattc
     4621 agaagaaaga tggcggctat ctctacacca ccactgatat cgcctgtgcg aaatatcgtt
     4681 atgaaacact gcatgccgat cgcgtgctgt attacatcga ctcccgtcag catcaacacc
     4741 tgatgcaggc atgggcgatc gtccgtaaag caggctatgt accggaatcc gtaccgctgg
     4801 aacaccacat gttcggcatg atgctgggta aagacggcaa accgttcaaa acccgcgcgg
     4861 gtggtacagt gaaactggcc gatctgctgg atgaagccct ggaacgtgca cgccgtctgg
     4921 tggcagaaaa gaacccggat atgccagccg acgagctgga aaaactggct aacgcggttg
     4981 gtattggtgc ggtgaaatat gcggatctct ccaaaaaccg caccacggac tacatcttcg
     5041 actgggacaa catgctggcg tttgagggta ataccgcgcc atacatgcag tatgcataca
     5101 cgcgtgtatt gtccgtgttc cgtaaagcag aaattgacga agagcaactg gctgcagctc
     5161 cggttatcat ccgtgaagat cgtgaagcgc aactggcagc tcgcctgctg cagtttgaag
     5221 aaaccctcac cgtggttgcc cgtgaaggca cgccgcatgt aatgtgtgct tacctgtacg
     5281 atctggccgg tctgttctct ggcttctacg agcactgccc gatcctcagc gcagaaaacg
     5341 aagaagtgcg taacagccgt ctaaaactgg cacaactgac ggcgaagacg ctgaagctgg
     5401 gtctggatac gctgggtatt gagactgtag agcgtatgta agatccggct gctaacaaag
     5461 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     5521 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     5581 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5641 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5701 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5761 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5821 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5881 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5941 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6001 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     6061 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6121 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     6181 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     6241 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     6301 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     6361 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     6421 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     6481 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     6541 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     6601 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     6661 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     6721 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     6781 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     6841 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     6901 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     6961 tgtattactg tttatgtaag cagacagttt tattgt
//
