LOCUS       sequence-354275-        6812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier Q2LWG4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6812)
  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 6812)
  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: Q2LWG4
FEATURES             Location/Qualifiers
     source          1..6812
                     /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             1532..1561
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1614..1632)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1628..1675
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1712..2167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1799..1818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2009..2030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2193..2297
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2298..2366,2367..3158)
                     /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(2516..2535)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3329..3917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3818..3837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4071..4088
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4103..4242
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4168..4187)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4328..4350
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4344..4535)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5344..6426)
                     /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     6388..6407
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6427..6504)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6710..6729
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gccagacggt catagaagag agaccggtta
      121 cggagttatg aaaatggaga acattacgca agaactcgct gaaattgttg gtaaaaccta
      181 cgtatcctgg caggccgagg aaaggtggtt ctatggtagg gattctgggc tgatgcctgc
      241 acatcaacca gattatgttg tcttaccaag aaccgtcgag gaaattcagg ctattgtgag
      301 atttgctgga cgtgaggaca tacctgtcgt gcccatgggt gccggaatgt ctctaagcgg
      361 attggttatt ccccaaaagg gtggtattgt tgtggatatg aagcgtatgg accgaatttt
      421 gaaggttaat gaaactgcca gatatgctgt tgtggaaggc gggacttcca caggcgcatt
      481 gcatgcccac cttaaccgat attaccccag attacgtttt agtattcctg actccccagc
      541 aacaacgacg attgctgcta atgtcatgat tcacggccag ggtagactga ctcaacaata
      601 tggatttaac agcgatatgg tgtcaggcct tgaggttgtg ttaccgtcag gtgaactttg
      661 cagaatcggt tcctgtgcat tatcagatga ctggttttcc aaaggtccac ccttgcctga
      721 tctttctggt ctattcctgg gttggtttgg agctacaggt atcttgacta aggttggctt
      781 gaagctatac cctcgtaaga aaatgcgtga tgtagagatt ttcatcactg atagcccaga
      841 tttcgtacct ggaatactat atgaaattac acatacagag atggccgaag acatcaacat
      901 ctttgctcaa cctctacctc ttatcttcaa agacaatcac catattgtag tgtacttcac
      961 tggtgaggat cttgaggaac tagactttaa gaagaagaaa atctttcacg ctctagacgg
     1021 gttcataagg aataaagacg gtggattcat gacgatagga gaaggtatga gaaagaccct
     1081 attggatatg ccccagagaa gtgctactac atttgcagac gttacaagag gcggcgggtt
     1141 tgagtattcc ggtccaatta ttccgattga tcgttaccct caatgttctc gtaaattaga
     1201 cgaactggcc gttaaatacg atctgaaata ccatgctact gctagggtta taggtagagg
     1261 tcatgccatg atgttcgcct tctccttcac gtttaacagg gcagacgttg atatgatgga
     1321 gcgtactagg aaagctctgc atgaagcttc agaatacgcc ttatcaatag gtggactttt
     1381 ctggaaacca acactcgacg agcagagaat tgctcttgct aagatggacc cccaaactgc
     1441 taaactaatg aaaatgatca aggagaactt ggatccacag ggaataatga atcctggtaa
     1501 ctgggaagtt gctgaggctg ctgctcttga gcaccaccat caccaccacc accaccacca
     1561 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1621 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1681 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     1741 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1801 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1861 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1921 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1981 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2041 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2101 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2161 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2221 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2281 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2341 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2401 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2461 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2521 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2581 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2641 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2701 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2761 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2821 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2881 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2941 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3001 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3061 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3121 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3181 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3961 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4021 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4081 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4141 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4201 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4261 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4321 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4381 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4441 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4501 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4561 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4621 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4681 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4741 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4801 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4861 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4921 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     4981 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     5041 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5101 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5161 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5221 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5281 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5461 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5521 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5581 ataacatgag ctgtcttcgg tatcatcgta tcccactacc gagatatccg caccaacgcg
     5641 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5701 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5761 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5821 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     5881 gatttgctgg tgacccaatg cgaccagatg ctccactccc agtcgcgtac cgtcttcatg
     5941 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6001 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6061 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6121 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6181 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6241 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6301 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6361 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6421 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6481 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6541 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6601 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6661 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6721 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6781 cgtagaggat cgagatctcg atcccgcgaa at
//
