LOCUS       sequence-354374-        6599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier Q0IDM6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6599)
  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 6599)
  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: Q0IDM6
FEATURES             Location/Qualifiers
     source          1..6599
                     /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             1319..1348
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1401..1419)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1415..1462
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1499..1954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1586..1605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1796..1817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1980..2084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2085..2153,2154..2945)
                     /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(2303..2322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3116..3704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3605..3624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3858..3875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3890..4029
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3955..3974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4115..4137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4131..4322)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5131..6213)
                     /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     6175..6194
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6214..6291)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6497..6516
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtcgtccacg gccgttctgg ctccgaatag
      121 cgtcgaagaa ttaggtcaat taatcgcaga tttacacgct gaaggtcgtc cttgggttcc
      181 gtcagggatg ggatcgcgct tacattgggg tccgcctctg gccgcagatt ccggaccagt
      241 tgtttcgatg cgtcacttga gccgtattgt ggaccatgcc gttgatgatt taaccattac
      301 tgttgatgcc gggatgccgt tggctgattt gcaaacagcg ctggcggaac accaacaatg
      361 gctgccatta aactggccgt ggggctctaa tatggcgagc ccacagtccg cgggcagtat
      421 tggtgggttg gttgctcgcg gtttatcggg tgggttacgt caacgccacc tcggggtacg
      481 cgatcaaatt attggaatct ccctgatccg ctccgatggc gtgtctgcac gcgccggcgg
      541 ccgcgttgtt aagaatgttg ctggttatga tcttatgcgt ttgttatgtg ggagttgggg
      601 ttcattagca ttaattacag gcctgacgct gcgtgtgcaa ccgattcgcg agcctcgcgg
      661 gcaactggtg ctggaaggtg aattaaacca gcttgaggcg tttcgcgcag aggttgtcgg
      721 tagttctttt accccggagc gtattgattg ggaattacgt cctgatcaag gtccccgtat
      781 cctgcttggg gtggcttcta ttagtgatgg cgcggtagtt gatcagctga atcgcttaga
      841 gaccattgcc gcaaagcaag gggtgagctc tgaacgcatg ttatgggaga atctcattcc
      901 ggaaggttcc gccactagcg aagatccgcc gtggctgtta cgtctttgtt taccgccagc
      961 ggagttacag catttgttct cgagccgtga gtgtaccgcc ctgaccggtt ggcattggca
     1021 attagcggcc ggagcaggct caggagaagc atggcaacat tcgggcgcac ctactccgga
     1081 gtacttgatt gaggcattac gtcgtcgcgt tattgcatta ggtggtcagt tgtctgtcct
     1141 gattcaaccg ggcgcccaac cgggtgctct ccctgcatgg ttagatgctc ccagccgtcc
     1201 gctgattgaa gcggtgaagc gccagtttga tccgaaacaa cagctgagcc gtggtcgctt
     1261 gccgggcgtt gctgcgccat tgaatagcgc acattcggga aatgcggccg cactcgagca
     1321 tcatcaccat caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     1381 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1441 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     1501 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1561 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1621 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1681 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1741 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1801 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1861 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1921 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1981 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2041 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2101 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2161 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2221 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2281 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2341 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2401 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2461 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2521 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2581 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     2641 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2701 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2761 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2821 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2881 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2941 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3001 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3061 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3241 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     3901 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     3961 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     4021 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4081 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4141 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     4201 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4261 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4321 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4381 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4441 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4501 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4561 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4621 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4681 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     4741 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     4801 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     4861 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     4921 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     4981 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     5041 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     5101 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5161 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5221 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5281 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5341 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     5401 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5461 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5521 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5581 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     5641 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     5701 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     5761 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     5821 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     5881 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     5941 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     6001 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     6061 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     6121 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     6181 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6241 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6301 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     6361 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6421 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6481 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     6541 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaat
//
