LOCUS       sequence-354298-        6476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier Q92NB8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  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 6476)
  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: Q92NB8
FEATURES             Location/Qualifiers
     source          1..6476
                     /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             1196..1225
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1278..1296)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1292..1339
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1376..1831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1463..1482)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1673..1694
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1857..1961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1962..2030,2031..2822)
                     /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(2180..2199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2993..3581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3482..3501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3735..3752
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3767..3906
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3832..3851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3992..4014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4008..4199)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5008..6090)
                     /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     6052..6071
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6091..6168)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6374..6393
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gagtaaagga gaagaagcca catgtccgcc
      121 ccacctggga gacttcgagt ctttgcaaga aattatccag aaagcgcaag gggccttacc
      181 gaaagaaaag tgggacagct tggtaggtgg ggccgaaaca gagacgacgt taaaacgtaa
      241 tcgtttagcc attgactcaa ttgcgttcaa gcctcgcgtt ctgcgtaacg tgagcgtggt
      301 ggatctgtcg attgagcatt ttggtcgtcg cttacgtctg ccgatctttc tcgcgcccac
      361 aggcccatta aatttatttg gtccgggcgg tggcgcagct gtggcaagtg gggcgcaggt
      421 atttggagtt gcccacatgc tgagtagcgg ttgtaccccg ctggagagcg tagccgaagc
      481 tgccccgtcc gctctgcgta tggcacagtt gtacgtgcgc ggcgatgacg cgtctgtaca
      541 caagtatgtg gggcgtgcct tggcttccgg gtgcgcagct atttgtctga ccgtggattc
      601 ggctgtactg gcacgtcgtg atcgcgatat tgcaaatcgc caccgtacag ccggcttagg
      661 caagtggccg ggtcaagcgt atcaagcggg gctggattgg cgtacggtta agctcatcaa
      721 agattcttat gatattccgt tagtgttgaa aggtattgca accgtagagg atgcgcgtat
      781 tgcggtggac cacggtgtag actggatcta tgtgtcgaat catggcggcc gccaacttga
      841 tcatggtcgc ggtaccatgg atgttctgcc cgaaatcatc gatgccgttg gcggccaggc
      901 gaaagtaatg gttgatggtg gcttctgtcg tggcacggat atcattaaag cgctggcgat
      961 tggtgcgaat ctggttggtc tcggtcgcat gcaatgttac gcattagctg caggcggcga
     1021 agcagccatc atccgcatgc tggaattaat cgaagatgag atgcttcgct ccatggcatt
     1081 acttggcgtt ccaacaatcg gcgacctgga tcgctcctac ctgtaccccg cagttccggt
     1141 tacgatccca agtgcgttat ctgcgtttcc gttaatcaac gcggccgcac tcgagcatca
     1201 tcaccatcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     1261 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1321 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     1381 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1441 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1501 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1561 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1621 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1681 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1741 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1801 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     1861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1921 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2341 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     2521 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2701 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2821 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3121 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3661 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3721 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3781 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     3841 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3901 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3961 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4021 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     4081 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     4141 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     4201 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     4261 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     4321 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     4381 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     4441 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     4501 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     4561 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     4621 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     4681 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     4741 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     4801 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     4861 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     4921 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     4981 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5041 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5101 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     5161 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     5221 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     5281 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     5341 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     5401 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     5461 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     5521 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     5581 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     5641 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     5701 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     5761 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     5821 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     5881 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     5941 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     6001 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     6061 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     6121 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     6181 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     6241 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     6301 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     6361 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6421 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaat
//
