LOCUS       sequence-354364-        6872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier B8MKR3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6872)
  AUTHORS   Rembeza E, Engqvist MKM
  TITLE     Experimental and computational investigation of enzyme functional 
            annotations uncovers misannotation in the EC 1.1.3.15 enzyme class.
  JOURNAL   PLoS Comput Biol. 2021 Sep 23;17(9):e1009446. doi: 
            10.1371/journal.pcbi.1009446. eCollection 2021 Sep.
  PUBMED    34555022
REFERENCE   2  (bases 1 to 6872)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Comput
            Biol."; date: "2021-09-23"; volume: "17(9):e1009446. doi"; pages: " 
            10.1371/journal.pcbi.1009446. eCollection 2021 Sep"
COMMENT     Sequence Label: B8MKR3
FEATURES             Location/Qualifiers
     source          1..6872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1592..1621
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1674..1692)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1688..1735
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1772..2227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1859..1878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2069..2090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2253..2357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2358..2426,2427..3218)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2576..2595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3389..3977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3878..3897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4131..4148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4163..4302
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4228..4247)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4388..4410
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4404..4595)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5404..6486)
                     /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"
     primer_bind     6448..6467
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6487..6564)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6770..6789
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggcacgtgta ctcgatgctg ccgaagtagc
      121 caaacacaat actcccgagt cgtgctgggt tattttgtat ggaaaggtgt acgatgtcac
      181 ggagtttatc agctcacatc ctggcggcgt gaaagtcatc ttgcgtctcg ccggcacgga
      241 tgcgacggaa gagtacgacc cgattcatcc gccgggcacc ctcgaggaaa atttaaaacc
      301 ggaagcactg ctgggcacag tggatcccga gactcttcct aagccagata aacagcagtc
      361 gagcccggaa gaggaagagc agggtccgcc gccaatggag actctgttta atctggatga
      421 gattgaacaa gtcgcgacca agcaggttag ccgcaaggcc tggggctact actactcagc
      481 cgcagatgac ttgatcagta agaacttcaa tcgcgagatc taccgtagca tcctgctgcg
      541 cccgcgtgtg ttcatcgacg ttggcaaatg tgacttaagc acgactattc tgggccatcg
      601 tgttggtctg cccatttata tttcgccggc agccatggcc cgcctggcac accccgcggg
      661 tgaggccgga atcgcagcgg cttgtcgtgg ctttggggca atgcaaatga tcagtaataa
      721 tgcgtcaatg agccctgaac aaattgtcga gaacgccgcg cctgaccaag ttttcggctg
      781 gcaactttac gtgcagatgg aacgtaagaa aagcgaggcc atgttggcgc gtgtggaaaa
      841 gctgaaagct atcaagtgtg tcatcctgac tttggatgca cctgtacctg gcaagcgtga
      901 ggatgatatg cgcacagata atattggcaa gaagttaccc gtgtcaagcg ccaaagtcgc
      961 tgaaaaggaa gtagagacac ttccggatgg cacgccggtt cccaccgatg gcggtggtgg
     1021 cgtaggcaag caacttttcg cgggaacagc ctatgatctg acttggaagg aaactttgac
     1081 ttggttgacc aaggtgacca agttgcctat catcctgaag gggctccaaa cacatgagga
     1141 cgcctatatt gcgagtctgt acgcgcctca agttaaagga atcatcctga gcaatcacgg
     1201 cggccgtgcg ttagatactg ctccgccggc tgtccatacc cttttagaaa tccgcaaata
     1261 ttgccctgag gtctttgata agattgaggt gctggttgat ggtggaatcc gtcgtggtac
     1321 agacgtggtt aaagccctgt gcctcggggc ccgtgcggtt gggattggtc gtcctgcgct
     1381 gtggggcctt ggggccggcg ggatcgcagg cgtggaacgt acgcttgaaa tcctggcgga
     1441 cgagacgaag acatgtatgc aactgctggg cgtagagaaa attagtgatc tcggtccgga
     1501 gtacatcaat agtcgcattg ttgaacagca gatttacgat ggtccctctg gactggaaaa
     1561 ggcgcgcgct aagttagcgg ccgcactcga gcatcatcac catcaccacc accaccacca
     1621 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1681 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1741 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     1801 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1861 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1921 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1981 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2041 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2101 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2161 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2221 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2281 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2341 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2401 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2461 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2521 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2581 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2641 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2701 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2761 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2821 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2881 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2941 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3001 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3061 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3121 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3181 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3241 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3301 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3361 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3421 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3481 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3541 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3601 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3661 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3721 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3781 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3841 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3901 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3961 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4021 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4081 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4141 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4201 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4261 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4321 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4381 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4441 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4501 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4561 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4621 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4681 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4741 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4801 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4861 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4921 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4981 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     5041 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     5101 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5161 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5221 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5281 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5341 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5401 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5461 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5521 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5581 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5641 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     5701 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5761 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5821 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5881 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     5941 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     6001 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6061 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6121 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6181 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6241 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6301 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6361 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6421 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6481 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6541 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6601 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6661 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6721 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6781 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6841 cgtagaggat cgagatctcg atcccgcgaa at
//
