LOCUS       sequence-354462-        6692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier Q5WFZ3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  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 6692)
  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: Q5WFZ3
FEATURES             Location/Qualifiers
     source          1..6692
                     /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             1412..1441
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1494..1512)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1508..1555
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1592..2047
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1679..1698)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1889..1910
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2073..2177
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2178..2246,2247..3038)
                     /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(2396..2415)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3209..3797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3698..3717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3951..3968
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3983..4122
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4048..4067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4208..4230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4224..4415)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5224..6306)
                     /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     6268..6287
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6307..6384)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6590..6609
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtcagatctg aaaagcgcgt tcccggaatt
      121 ggcgatggag ccaggcagcc cggatagctg gctcggtaat ggcggtcatg tgcacgtata
      181 ccccaccagc gagaaagagg tcgtcggcct gttaaagttt gcttatgatc acgggaagag
      241 tgtgagtgtg caaggaaacg ggagtaaacg tggcttcggc ggattaaagg aatcctacga
      301 cgtgctgctg tcactgagta aatatacggg agtgattgag cacagtccag gtgatatggt
      361 cgtaacagtt aaagctggga cggcgttcca cgagttgcaa gcgtatctgg caaagcatca
      421 acaacaggtc agtctcgacc cggctatgcc gcatcttagc accgtgggcg gtgtaatcgc
      481 ttccaatgac agcgggccga agcgcatggg atatggttcg gctcgcgata ttgttcttgg
      541 tctgaaagtg gtatatccgg atgggacggt gatccgcacg ggcgggaagg ttgtgaagaa
      601 tgtggcgggc tatgacatga acaagctgtt cattggttct atgggtacga ttggcgttat
      661 tacggaaatc accctgaagc tgatgccgct gccgaaatgt gagagccttg ttttcattcc
      721 atttcaagac aaccgtttag acgaacttcg tagctttgta gtgacccaat ttgactcaac
      781 cttagaaccc accgcgtttg agttcatctc tccaagcctt gccgagcaat tgaccggtcg
      841 caagcagtgg gcgctggcta ttggtctgga agatgttgaa tcttctgtta tctaccaaga
      901 aaacatcgta aagcaagcag ttccgaaaga ggcggccgat gtgcacatct tccgctcgga
      961 agaggcgcgc gcgttttgga atggccttat gaatatctcc ccgaacggca tgggcgaacc
     1021 gagtgaacaa gctgccatta aagtcggcct caaaattggt gtcaagaata ttgatgtcct
     1081 gcaaatgctt gccgagtgcg tagcccttca gcagcgtcac aacttggtgg tttatggtca
     1141 tggtgggttc ggccacggcc tctgccatgt tgttcttcaa ggaacaagtg aagacgtaga
     1201 agcggcgatt caggccctgc gtggcacggc gaagaaactt aacgggtatg ttatcgtaaa
     1261 acatatccca ttcttcctgc gccagagcat taatgtttgg ggcgataagc cggattattt
     1321 cttcctgttc gagggcatta aagcgagcat cgatccacgc cgtgttctga atgaaacccg
     1381 ctttatcggt ggcattgcgg ccgcactcga gcatcatcac catcaccacc accaccacca
     1441 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1501 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1561 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     1621 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1681 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1741 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1801 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1861 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1921 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1981 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2041 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2101 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2161 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2221 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2281 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2341 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2401 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2461 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2521 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2581 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2641 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2701 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2761 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2821 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2881 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2941 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3001 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3061 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3121 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3181 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3241 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3301 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3361 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3421 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3481 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3541 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3601 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3661 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3721 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3781 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3841 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3901 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3961 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4021 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4081 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4141 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4201 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4261 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4321 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4381 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4441 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4501 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4561 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4621 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4681 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4741 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4801 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     4861 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     4921 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     4981 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5041 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5101 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5161 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5221 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5281 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5341 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5401 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5461 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     5521 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5581 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5641 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5701 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     5761 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     5821 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     5881 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     5941 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6001 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6061 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6121 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6181 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6241 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6301 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6361 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6421 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6481 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6541 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6601 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6661 cgtagaggat cgagatctcg atcccgcgaa at
//
