LOCUS       sequence-354359-        6572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0U6K8E5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  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 6572)
  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: A0A0U6K8E5
FEATURES             Location/Qualifiers
     source          1..6572
                     /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             1292..1321
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1374..1392)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1388..1435
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1472..1927
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1559..1578)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1769..1790
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1953..2057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2058..2126,2127..2918)
                     /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(2276..2295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3089..3677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3578..3597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3831..3848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3863..4002
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3928..3947)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4088..4110
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4104..4295)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5104..6186)
                     /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     6148..6167
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6187..6264)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6470..6489
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtacgacgcg atcgtgattg gtgctggtct
      121 tgttggtttg ggtgtagtaa acgcgctcca agaagcggac ccgcaacgcc gcattcttat
      181 tttagagaaa gagcgtggcc cggcggctca tcagtcgggc cacaacagca acgtggtaca
      241 ttcgggaatt tactatgccc caggtagtct gaaagcccag ctggcgaagc aaggtaaccg
      301 tagtatgttc gcgttttgtc gcgaacacga tttataccac gatcagtgcg gaaaagtgat
      361 tgtggcgacg caactgaacg agctcgatcg tcttgagaac atttaccagc gcgggctgca
      421 gaacggcctg caggtaactc gcctgtcaca ggcccaatta cgtgagcgcg agccgcacgt
      481 taatgggttg gaagcgctgc tggtgccgga tgccggcatc gtcaactacg ccgaagttgc
      541 ggccaaactg gctgaaatca tcgttcgccg cggcggtgaa atcgcttatg gccatcaggt
      601 agaggcgatc catgaacatg cggcgggcgt caccgttact gccgcgggcg atagctttga
      661 agggaaactg ttaatcaact gtgccggatt gttttcagat cgcattgcga aacttgcggg
      721 gtatgaaacg gggatgaaaa ttgtgccgtt ccgcggcgag tattacgttt tagatgatgc
      781 aaagaaccat ctggttaacc accttatcta tccagtaccg aaccccgatt tcccgttcct
      841 gggagtgcat tttacccgca tgtataacgg taaacgcgac gttggaccga atgcagtcct
      901 ggcgttcaag cgcgagggct atcgcaagtg tgactttagt ctgcgtgatt tgtctgaggt
      961 attaggttac cgtggctttt ggaaaatcgc gggaaattac ctgggtgaag gtctcgcaga
     1021 gatgcgccgt agtcttagcc gccgtcgttt cgccgaaaat gcccgccgcc tgattccggc
     1081 actgcaagag caagatattc gtccgggccc ggccggtgtg cgtgcacaag ctcttacggc
     1141 ggacggtaaa ctggtagacg actttcactt tgtgcaagga cagcgtagct tacatgtgtg
     1201 taacgcaccg tcgccggcag cgactgcatc cctggaaatc ggtcgtgaga tcgtccgccg
     1261 ccacctgagt gcgtttgcgg ccgcactcga gcatcatcac catcaccacc accaccacca
     1321 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1381 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1441 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     1501 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1561 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1621 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1681 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1741 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1801 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1861 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     1921 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2041 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2161 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2221 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2281 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2341 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2461 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2521 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2581 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2641 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2701 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2761 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2821 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2881 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2941 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3001 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3061 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3121 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3181 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3241 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3301 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3361 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3421 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3481 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3541 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3601 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3661 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3721 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3781 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3841 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3901 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3961 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4021 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4081 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4141 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4201 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4261 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4321 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4381 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4441 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4501 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4561 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4621 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4681 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     4741 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     4801 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     4861 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     4921 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     4981 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5041 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5101 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5161 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5221 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5281 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5341 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     5401 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5461 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5521 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5581 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     5641 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     5701 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     5761 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     5821 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     5881 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     5941 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6001 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6061 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6121 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6181 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6241 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6301 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6361 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6421 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6481 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6541 cgtagaggat cgagatctcg atcccgcgaa at
//
