LOCUS       sequence-354402-        6899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier Q0I8G2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6899)
  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 6899)
  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: Q0I8G2
FEATURES             Location/Qualifiers
     source          1..6899
                     /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             1619..1648
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1701..1719)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1715..1762
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1799..2254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1886..1905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2096..2117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2280..2384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2385..2453,2454..3245)
                     /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(2603..2622)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3416..4004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3905..3924
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4158..4175
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4190..4329
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4255..4274)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4415..4437
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4431..4622)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5431..6513)
                     /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     6475..6494
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6514..6591)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6797..6816
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gatgacgaaa cttcctaagc tgcaattatt
      121 tggcacttca tcagtcaatt atgactggcc ggcgttagaa cgcgagctgc gcggtttcct
      181 tccgtcgaag agtgtagtag cgaagcgtca ggaattatta tcatatgatt gcgatggcct
      241 gacgatggat cgtcacatgc ctcgcttagc agttctgcct gaaacagccg agcaagtggc
      301 acaaattttg gcatgttgtc atcgtctggg catccccttt gtcgcgcgtg gttcaggtac
      361 agggctttcg ggcggagcac tggtggaaca agaagccctg ctggtcgtta cctctcgtat
      421 gcgtcgcatc ttggacattg atctggacaa tcagaccatt accgtggagc cgggtgtgat
      481 caatagttgg gtttctcgtg cggtggtcgg tgatgggttc tattatgccc cagatccgtc
      541 ttcccaagtt gtttgcagca tcggtgggaa cgtcgctgaa aacagcggtg gcgtgcactg
      601 tttaaaatac ggtgttacca gtaatcacgt actgagcctg gacgtggtgc tgcccaacgg
      661 aacgccgact cgccttggta cctctctgtg tgacgctgcc gaactggacc tgcgcggggt
      721 gtttattggt tcagagggca ccctgggcat cgctactgcc attactctgc gtcttcttcg
      781 cgctcctgac gccgtgggcg tcttacttgc cgactttccg agtatgcaag ccgcgggcga
      841 agcggtgcgt ctgatcaccc gtgccggtgt acttcctgca ggtttagaga ttatggacca
      901 gacctgtatc caggccgtta acgaagcctt tggtgaagaa gagtacccac ccgaggcagg
      961 ggccgtcttg ctgatcgaac tggatggcca agagcaagaa gtaaaacagg cagttactgt
     1021 gaccaccgct ttatgtcgtg aagccggtgc cggggcaatt cgcgaagcgt ggagcgagga
     1081 agatcgcgca cgcttgtgga agggccgtaa atcggccatt agtgcgctgg gtcgccagta
     1141 cccgtcatat tacctgcagg atggcgttgt tcctcgtacc gccttgccac gcgttctgga
     1201 agcgattgac cagctttcag cggaacacgg cttagtagtt gcgaatgtct tccacgccgg
     1261 cgatggtaac cttcatcccc tgattctgta tcgttcgtcc gaacagggtg tgaatgagcg
     1321 cgtcaaggcc ttgggcggtg caattatgaa cttatgtttg gaagtaggtg gctccatctc
     1381 aggtgagcat ggagtgggaa gcgacaagcg ttgttacctc gatcagatgt tcagcgcgga
     1441 agatctgatc tcaatgcagt gggtccgtct ggcgtttgac cccttgggtc gtgcgaaccc
     1501 cggcaagatc ttcccaaccc ctaagagctg cggtgagagc ttacgtcgca gtgtacaatt
     1561 gcaagcagag ggacagagcc tgccggaaga agcaattgtg tatgcggccg cactcgagca
     1621 tcatcaccat caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     1681 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1741 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     1801 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1861 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1921 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1981 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2041 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2101 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2161 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2221 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     2281 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2341 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2401 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2461 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2521 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2581 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2641 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2701 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2761 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2821 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2881 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     2941 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3001 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3061 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3121 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3181 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3241 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3301 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3361 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3421 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3481 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3541 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3601 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3661 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3721 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3781 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3841 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3901 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3961 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4021 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4081 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4141 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4201 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     4261 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     4321 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4381 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4441 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     4501 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4561 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4621 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4681 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4741 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4801 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4861 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4921 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4981 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     5041 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     5101 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     5161 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     5221 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     5281 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     5341 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     5401 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5461 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5521 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5581 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5641 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     5701 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5761 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5821 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5881 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     5941 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     6001 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     6061 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     6121 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     6181 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     6241 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     6301 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     6361 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     6421 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     6481 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6541 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6601 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     6661 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6721 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6781 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     6841 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaat
//
