LOCUS       pET28a-ValRS            8099 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Nucleus PURE: pET28a-ValRS in pET28a protein expression plasmid. See
            https://nucleus.bnext.bio/ for more details..
ACCESSION   pET28a-ValRS
VERSION     pET28a-ValRS
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..6501
                     /codon_start=1
                     /standard_name="ValRS"
                     /label="ValRS"
                     /translation="MEKTYNPQDIEQPLYEHWEKQGYFKPNGDESQESFCIMIPPPNVT
                     GSLHMGHAFQQTIMDTMIRYQRMQGKNTLWQVGTDHAGIATQMVVERKIAAEEGKTRHD
                     YGREAFIDKIWEWKAESGGTITRQMRRLGNSVDWERERFTMDEGLSNAVKEVFVRLYKE
                     DLIYRGKRLVNWDPKLRTAISDLEVENRESKGSMWHIRYPLADGAKTADGKDYLVVATT
                     RPETLLGDTGVAVNPEDPRYKDLIGKYVILPLVNRRIPIVGDEHADMEKGTGCVKITPA
                     HDFNDYEVGKRHALPMINILTFDGDIRESAQVFDTKGNESDVYSSEIPAEFQKLERFAA
                     RKAVVAAVDALGLLEEIKPHDLTVPYGDRGGVVIEPMLTDQWYVRADVLAKPAVEAVEN
                     GDIQFVPKQYENMYFSWMRDIQDWCISRQLWWGHRIPAWYDEAGNVYVGRNEDEVRKEN
                     NLGADVVLRQDEDVLDTWFSSALWTFSTLGWPENTDALRQFHPTSVMVSGFDIIFFWIA
                     RMIMMTMHFIKDENGKPQVPFHTVYMTGLIRDDEGQKMSKSKGNVIDPLDMVDGISLPE
                     LLEKRTGNMMQPQLADKIRKRTEKQFPNGIEPHGTDALRFTLAALASTGRDINWDMKRL
                     EGYRNFCNKLWNASRFVLMNTEGQDCGFNGGEMTLSLADRWILAEFNQTIKAYREALDS
                     FRFDIAAGILYEFTWNQFCDWYLELTKPVMNGGTEAELRGTRHTLVTVLEGLLRLAHPI
                     IPFITETIWQRVKVLCGITADTIMLQPFPQYDASQVDEAALADTEWLKQAIVAVRNIRA
                     EMNIAPGKPLELLLRGCSADAERRVNENRGFLQTLARLESITVLPADDKGPVSVTKIID
                     GAELLIPMAGLINKEDELARLAKEVAKIEGEISRIENKLANEGFVARAPEAVIAKEREK
                     LEGYAEAKAKLIEQQAVIAAL"
     CDS             6502..6516
                     /codon_start=1
                     /label="Linker"
                     /translation="GGGGS"
     CDS             6517..6534
                     /codon_start=1
                     /label="His6"
                     /translation="HHHHHH"
     terminator      6608..6655
                     /label="T7 terminator"
                     /note="Color: #ffffff NCBI Feature Key: terminator
                     Transferred From: T7_terminator Transferred Similarity:
                     100.00%"
     rep_origin      6692..7147
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(7239..8054)
                     /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 ggagaaaaca
     3661 tacaaccctc aggacattga acagccgctg tatgagcatt gggaaaaaca gggttacttc
     3721 aaaccgaacg gcgacgaaag ccaggagtcc ttttgcatca tgattcctcc accgaacgtg
     3781 accggtagcc tgcacatggg ccatgccttc caacagacca tcatggatac catgattcgt
     3841 taccaacgca tgcagggcaa aaacaccctg tggcaagtcg ggactgacca cgcgggaatt
     3901 gccacacaga tggttgttga acggaagatt gcggccgaag agggtaagac ccgccatgac
     3961 tatggtcgtg aggcctttat cgataaaatt tgggaatgga aagcggaatc aggtggcact
     4021 atcacccgtc agatgcgccg tctgggtaac tcagtcgatt gggaacgtga acgcttcact
     4081 atggacgaag gtttaagcaa cgctgtgaaa gaggtgtttg tccgtttgta caaagaagat
     4141 ttgatctacc ggggcaaacg ccttgttaac tgggatccga aactgcggac ggctatttct
     4201 gatttagagg tcgaaaaccg cgagtccaag ggtagcatgt ggcacatccg gtatccttta
     4261 gcagatggcg cgaaaactgc ggatggaaaa gattatctgg ttgtagcaac aacccgccca
     4321 gaaacactgc tgggtgacac cggcgtggcc gtgaacccgg aagatcctcg ttataaggat
     4381 ttgattggca agtatgtgat cttgccgctg gttaatcgtc gtatccctat tgttggtgac
     4441 gaacatgcgg acatggagaa aggcaccggg tgcgtgaaaa tcacccctgc ccatgatttt
     4501 aacgattacg aggttggcaa acgccatgca ttacctatga ttaatatctt gacgttcgat
     4561 ggagacatcc gggaaagcgc ccaggttttt gatacgaagg gcaatgaaag cgacgtgtac
     4621 tcgtcggaaa tcccagcgga gttccaaaaa ttagaacgtt tcgccgcgcg caaagccgtg
     4681 gtggccgccg ttgatgcact gggcctgtta gaagagatca aaccccatga tcttacagta
     4741 ccgtacggcg atcgtggcgg ggtggtcatt gagccaatgt taacggacca gtggtatgtt
     4801 cgggccgacg ttctggcaaa gccggccgtg gaagcggtag aaaatggtga cattcaattc
     4861 gtcccgaaac aatatgaaaa catgtacttc tcgtggatgc gcgacattca agattggtgt
     4921 attagtcggc agttgtggtg gggccatcgt attccagcct ggtatgatga ggccggtaat
     4981 gtgtacgttg gccgtaatga ggatgaagtc cgtaaagaga acaatctggg ggcggatgtg
     5041 gtgctgcgcc aggatgagga tgtgcttgac acctggttct ccagtgccct ttggacattc
     5101 agcacgttgg gctggccaga aaacacagat gcgctgcgcc aatttcaccc tacatcagta
     5161 atggtgtccg ggttcgatat tatttttttc tggattgcgc gtatgatcat gatgaccatg
     5221 cattttatta aagatgaaaa cggtaagccg caggtcccct ttcacacagt atatatgact
     5281 ggattaattc gcgacgatga aggccaaaaa atgtccaaga gtaagggtaa tgtgattgat
     5341 ccactggata tggtggatgg tatctcgctg ccggagttgt tagaaaaacg cacaggcaat
     5401 atgatgcagc cacaacttgc cgataaaatc cgcaaacgta cggaaaaaca atttcctaat
     5461 ggcatcgagc cccacggtac ggacgcgctg cggttcaccc ttgccgcgtt agcgtccacg
     5521 ggacgcgata ttaattggga tatgaaacgc ttggaaggct accgcaactt ctgcaataaa
     5581 ctgtggaatg cttcacggtt cgtcctgatg aacacagaag gtcaagattg cggttttaat
     5641 ggtggggaaa tgacacttag cctggccgac cgttggatct tggcggaatt caatcaaacc
     5701 atcaaagcct atcgcgaggc gctggacagc tttcgtttcg acattgcggc tgggattctg
     5761 tatgaattta cttggaatca gttttgtgat tggtaccttg aattgaccaa accagtgatg
     5821 aacggtggca cagaggcgga gcttcgcggc acccgccata cgcttgtgac tgttttagaa
     5881 ggcctgctgc gcttagctca cccaattatc ccatttatta cagagacaat ctggcaacgt
     5941 gtgaaggttt tgtgtggtat tacggcggac accatcatgc tgcaaccgtt tccccagtac
     6001 gatgcgagtc aggtggacga agcagcgctg gcggacacag aatggcttaa acaagcaatt
     6061 gtcgcagtcc gtaacattcg tgcggaaatg aatattgcac cgggtaaacc tctggaactt
     6121 ctgttacggg gatgctcagc agacgccgaa cgtcgggtaa atgaaaatcg tggctttctg
     6181 cagacgttgg ctcgcttgga atcaatcacg gtattgcccg cggatgacaa gggaccggtg
     6241 tcagttacaa aaatcatcga tggagcggaa cttctgattc caatggccgg attaatcaat
     6301 aaagaagatg aactggcgcg tttagcaaaa gaggtagcaa aaattgaggg cgagatttct
     6361 cgcatcgaaa ataaattggc gaatgagggt tttgttgccc gggcaccaga agctgtcatt
     6421 gctaaagagc gcgagaaact ggaggggtat gcggaggcca aagcaaaact gattgagcag
     6481 caggctgtga ttgccgctct gggaggcggt ggatctcacc atcaccacca tcattgaaga
     6541 gcttgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     6601 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     6661 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     6721 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6781 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6841 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6901 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6961 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     7021 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     7081 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     7141 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7201 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     7261 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     7321 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     7381 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     7441 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     7501 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     7561 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     7621 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     7681 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     7741 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     7801 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     7861 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     7921 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     7981 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     8041 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgt
//
