LOCUS       pET28a-ProRS            6962 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Nucleus PURE: pET28a-ProRS in pET28a protein expression plasmid. See
            https://nucleus.bnext.bio/ for more details..
ACCESSION   pET28a-ProRS
VERSION     pET28a-ProRS
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..5364
                     /codon_start=1
                     /standard_name="ProRS"
                     /label="ProRS"
                     /translation="MRTSQYLLSTLKETPADAEVISHQLMLRAGMIRKLASGLYTWLPT
                     GVRVLKKVENIVREEMNNAGAIEVSMPVVQPADLWQESGRWEQYGPELLRFVDRGERPF
                     VLGPTHEEVITDLIRNELSSYKQLPLNFYQIQTKFRDEVRPRFGVMRSREFLMKDAYSF
                     HTSQESLQETYDAMYAAYSKIFSRMGLDFRAVQADTGSIGGSASHEFQVLAQSGEDDVV
                     FSDTSDYAANIELAEAIAPKEPRAAATQEMTLVDTPNAKTIAELVEQFNLPIEKTVKTL
                     LVKAVEGSSFPQVALLVRGDHELNEVKAEKLPQVASPLTFATEEEIRAVVKAGPGSLGP
                     VNMPIPVVIDRTVAAMSDFAAGANIDGKHYFGINWDRDVATPEVADIRNVVAGDPSPDG
                     QGRLLIKRGIEVGHIFQLGTKYSEALKASVQGEDGRNQILTMGCYGIGVTRVVAAAIEQ
                     NYDERGIVWPDAIAPFQVAILPMNMHKSFRVQELAEKLYSELRAQGIEVLLDDRKERPG
                     VMFADMELIGIPHTIVLGDRNLDNDDIEYKYRRNGEKQLIKTGDIVEYLVKQIKG"
     CDS             5365..5379
                     /codon_start=1
                     /label="Linker"
                     /translation="GGGGS"
     CDS             5380..5397
                     /codon_start=1
                     /label="His6"
                     /translation="HHHHHH"
     terminator      5471..5518
                     /label="T7 terminator"
                     /note="Color: #ffffff NCBI Feature Key: terminator
                     Transferred From: T7_terminator Transferred Similarity:
                     100.00%"
     rep_origin      5555..6010
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(6102..6917)
                     /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 gcgtaccagc
     3661 caatatcttt tgtcgaccct gaaggagact ccagcggatg ccgaggttat tagccaccag
     3721 ttgatgcttc gtgccggtat gattcgtaaa ctggcaagcg ggctgtatac atggctgccg
     3781 actggtgtac gtgttcttaa aaaagtcgaa aacatcgtcc gggaggagat gaacaacgcg
     3841 ggcgccattg aggtgagcat gcccgtggtc cagccggcgg atctttggca ggagagtggt
     3901 cggtgggaac agtatggccc ggaattgctg cgtttcgttg atcgtgggga acgcccgttt
     3961 gtgctggggc caacacatga ggaagtgatc accgatctga tccgcaacga actgtcaagc
     4021 tacaagcaat taccgttgaa tttctatcag atccagacca agttccgtga tgaagtccgt
     4081 ccgcgttttg gagtcatgcg gtcgcgcgag tttcttatga aagacgctta tagctttcat
     4141 acttcccagg agagcctgca ggaaacctat gatgcaatgt atgcagcgta ttctaaaatt
     4201 ttctcgcgta tgggtctgga ctttcgtgca gtacaggcgg acaccgggag catcggtggc
     4261 agcgcatccc acgaatttca agtgttggcg caatccggcg aagatgatgt tgtattcagc
     4321 gacaccagtg attatgcggc aaatatcgaa ctggcggagg caattgcccc taaggaaccg
     4381 cgtgccgccg caacccagga aatgacgctg gttgatacgc cgaatgcgaa aacgatcgca
     4441 gaactggtgg aacaatttaa tctgccaatt gaaaaaaccg ttaaaacgct gcttgttaaa
     4501 gcggtggaag ggtccagttt tccccaggtg gccttactgg ttcgcggaga tcatgaactg
     4561 aacgaagtga aagccgaaaa gctgccgcaa gtcgcatctc cccttacgtt tgctacagaa
     4621 gaggaaattc gtgcggtggt caaagcggga ccgggtagtc ttggtccagt gaatatgcca
     4681 attcccgtgg tgattgaccg taccgtcgcc gccatgagtg actttgcggc tggcgcgaac
     4741 atcgatggca agcattattt cggtatcaac tgggaccgcg acgtcgccac cccggaggtg
     4801 gcggatatcc gcaatgtagt cgcaggcgac ccatctccgg acggccaggg tcgcttgctt
     4861 atcaaacgtg ggatcgaagt tggccacatc ttccagttag gcacaaagta cagtgaagct
     4921 ctgaaagcat ctgtgcaggg tgaagatggc cgtaatcaaa ttctgactat gggctgctac
     4981 ggaatcggcg tgacgcgtgt tgtagcagca gcaatcgaac agaattatga tgaacgcggg
     5041 atcgtttggc cagatgcgat tgcccctttc caagtagcca ttttaccgat gaatatgcat
     5101 aaaagcttcc gtgtccagga acttgcggaa aagttatact ccgagctgcg tgcacagggt
     5161 attgaggttc ttcttgatga tcgcaaagaa cgcccgggcg ttatgtttgc ggacatggaa
     5221 ctgattggta tcccacatac gatcgttctg ggcgaccgga acttggataa cgatgacatt
     5281 gaatataaat atcgtcgcaa tggcgaaaaa cagctgatca agactggtga tatcgtcgaa
     5341 tatcttgtta agcagatcaa aggcggaggc ggtggatctc accatcacca ccatcattga
     5401 agagcttgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     5461 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     5521 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     5581 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5641 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5701 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5761 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5821 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5881 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5941 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     6001 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6061 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     6121 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     6181 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     6241 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     6301 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     6361 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     6421 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     6481 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     6541 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     6601 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     6661 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     6721 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     6781 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     6841 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     6901 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     6961 gt
//
