LOCUS       pET28a-LeuRS            7809 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Nucleus PURE: pET28a-LeuRS in pET28a protein expression plasmid. See
            https://nucleus.bnext.bio/ for more details..
ACCESSION   pET28a-LeuRS
VERSION     pET28a-LeuRS
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%"
     source          3648..6254
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             3648..6227
                     /codon_start=1
                     /label="LeuRS"
                     /translation="MQEQYRPEEIESKVQLHWDEKRTFEVTEDESKEKYYCLSMLPYPS
                     GRLHMGHVRNYTIGDVIARYQRMLGKNVLQPIGWDAFGLPAEGAAVKNNTAPAPWTYDN
                     IAYMKNQLKMLGFGYDWSRELATCTPEYYRWEQKFFTELYKKGLVYKKTSAVNWCPNDQ
                     TVLANEQVIDGCCWRCDTKVERKEIPQWFIKITAYADELLNDLDKLDHWPDTVKTMQRN
                     WIGRSEGVEITFNVNDYDNTLTVYTTRPDTFMGCTYLAVAAGHPLAQKAAENNPELAAF
                     IDECRNTKVAEAEMATMEKKGVDTGFKAVHPLTGEEIPVWAANFVLMEYGTGAVMAVPG
                     HDQRDYEFASKYGLNIKPVILAADGSEPDLSQQALTEKGVLFNSGEFNGLDHEAAFNAI
                     ADKLTAMGVGERKVNYRLRDWGVSRQRYWGAPIPMVTLEDGTVMPTPDDQLPVILPEDV
                     VMDGITSPIKADPEWAKTTVNGMPALRETDTFDTFMESSWYYARYTCPQYKEGMLDSEA
                     ANYWLPVDIYIGGIEHAIMHLLYFRFFHKLMRDAGMVNSDEPAKQLLCQGMVLADAFYY
                     VGENGERNWVSPVDAIVERDEKGRIVKAKDAAGHELVYTGMSKMSKSKNNGIDPQVMVE
                     RYGADTVRLFMMFASPADMTLEWQESGVEGANRFLKRVWKLVYEHTAKGDVAALNVDAL
                     TENQKALRRDVHKTIAKVTDDIGRRQTFNTAIAAIMELMNKLAKAPTDGEQDRALMQEA
                     LLAVVRMLNPFTPHICFTLWQELKGEGDIDNAPWPVADEKAMVEDSTLVVVQVNGKVRA
                     KITVPVDATEEQVRERAGQEHLVAKYLDGVTVRKVIYVPGKLLNLVVG"
     CDS             6228..6233
                     /codon_start=1
                     /label="Linker"
                     /translation="FE"
     CDS             6234..6251
                     /codon_start=1
                     /label="His6"
                     /translation="HHHHHH"
     terminator      6318..6365
                     /label="T7 terminator"
                     /note="Color: #ffffff NCBI Feature Key: terminator
                     Transferred From: T7_terminator Transferred Similarity:
                     100.00%"
     rep_origin      6402..6857
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(6949..7764)
                     /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 caagagcaat
     3661 accgcccgga agagatagaa tccaaagtac agcttcattg ggatgagaag cgcacatttg
     3721 aagtaaccga agacgagagc aaagagaagt attactgcct gtctatgctt ccctatcctt
     3781 ctggtcgact acacatgggc cacgtacgta actacaccat cggtgacgtg atcgcccgct
     3841 accagcgtat gctgggcaaa aacgtcctgc agccgatcgg ctgggacgcg tttggtctgc
     3901 ctgcggaagg cgcggcggtg aaaaacaaca ccgctccggc accgtggacg tacgacaaca
     3961 tcgcgtatat gaaaaaccag ctcaaaatgc tgggctttgg ttatgactgg agccgcgagc
     4021 tggcaacctg tacgccggaa tactaccgtt gggaacagaa attcttcacc gagctgtata
     4081 aaaaaggcct ggtatataag aagacttctg cggtcaactg gtgcccgaac gaccagaccg
     4141 tactggcgaa cgaacaagtt atcgacggct gctgctggcg ctgcgatacc aaagttgaac
     4201 gtaaagagat cccgcagtgg tttatcaaaa tcactgctta cgctgacgag ctgctcaacg
     4261 atctggataa actggatcac tggccagaca ccgttaaaac catgcagcgt aactggatcg
     4321 gtcgttccga aggcgtggag atcaccttca acgttaacga ctatgacaac acgctgaccg
     4381 tttacactac ccgcccggac acctttatgg gttgtaccta cctggcggta gctgcgggtc
     4441 atccgctggc gcagaaagcg gcggaaaata atcctgaact ggcggccttt attgacgaat
     4501 gccgtaacac caaagttgcc gaagctgaaa tggcgacgat ggagaaaaaa ggcgtcgata
     4561 ctggctttaa agcggttcac ccattaacgg gcgaagaaat tcccgtttgg gcagcaaact
     4621 tcgtattgat ggagtacggc acgggcgcag ttatggcggt accggggcac gaccagcgcg
     4681 actacgagtt tgcctctaaa tacggcctga acatcaaacc ggttatcctg gcagctgacg
     4741 gctctgagcc agatctttct cagcaagccc tgactgaaaa aggcgtgctg ttcaactctg
     4801 gcgagttcaa cggtcttgac catgaagcgg ccttcaacgc catcgccgat aaactgactg
     4861 cgatgggcgt tggcgagcgt aaagtgaact accgcctgcg cgactggggt gtttcccgtc
     4921 agcgttactg gggcgcgccg attccgatgg tgacgctgga agacggtacc gtaatgccga
     4981 ccccggacga ccagctgccg gtgatcctgc cggaagatgt ggtaatggac ggcattacca
     5041 gcccgattaa agcagatccg gagtgggcga aaactaccgt taacggtatg ccagcactgc
     5101 gtgaaaccga cactttcgac acctttatgg agtcctcctg gtactatgcg cgctacactt
     5161 gcccgcagta caaagaaggt atgctggatt ccgaagcggc taactactgg ctgccggtgg
     5221 atatctacat tggtggtatt gaacacgcca ttatgcacct gctctacttc cgcttcttcc
     5281 acaaactgat gcgtgatgca ggcatggtga actctgacga accagcgaaa cagttgctgt
     5341 gtcagggtat ggtgctggca gatgccttct actatgttgg cgaaaacggc gaacgtaact
     5401 gggtttcccc ggttgatgct atcgttgaac gtgacgagaa aggccgtatc gtgaaagcga
     5461 aagatgcggc aggccatgaa ctggtttata ccggcatgag caaaatgtcc aagtcgaaga
     5521 acaacggtat cgacccgcag gtgatggttg aacgttacgg cgcggacacc gttcgtctgt
     5581 ttatgatgtt tgcttctccg gctgatatga ctctcgaatg gcaggaatcc ggtgtggaag
     5641 gggctaaccg cttcctgaaa cgtgtctgga aactggttta cgagcacaca gcaaaaggtg
     5701 atgttgcggc actgaacgtt gatgcgctga ctgaaaatca gaaagcgctg cgtcgcgatg
     5761 tgcataaaac gatcgctaaa gtgaccgatg atatcggccg tcgtcagacc ttcaacaccg
     5821 caattgcggc gattatggag ctgatgaaca aactggcgaa agcaccaacc gatggcgagc
     5881 aggatcgcgc tctgatgcag gaagcactgc tggccgttgt ccgtatgctt aacccgttca
     5941 ccccgcacat ctgcttcacg ctgtggcagg aactgaaagg cgaaggcgat atcgacaacg
     6001 cgccgtggcc ggttgctgac gaaaaagcga tggtggaaga ctccacgctg gtcgtggtgc
     6061 aggttaacgg taaagtccgt gccaaaatca ccgttccggt ggacgcaacg gaagaacagg
     6121 ttcgcgaacg tgctggccag gaacatctgg tagcaaaata tcttgatggc gttactgtac
     6181 gtaaagtgat ttacgtacca ggtaaactcc tcaatctggt cgttggcttc gagcaccacc
     6241 accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     6301 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     6361 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
     6421 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6481 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6541 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6601 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6661 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6721 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6781 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6841 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6901 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
     6961 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     7021 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     7081 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     7141 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     7201 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     7261 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     7321 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     7381 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     7441 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     7501 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     7561 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     7621 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     7681 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     7741 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     7801 ttttattgt
//
