LOCUS       sequence-354453-        6404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier Q5FIR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  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 6404)
  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: Q5FIR1
FEATURES             Location/Qualifiers
     source          1..6404
                     /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             1124..1153
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1206..1224)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1220..1267
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1304..1759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1391..1410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1601..1622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1785..1889
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1890..1958,1959..2750)
                     /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(2108..2127)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2921..3509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3410..3429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3663..3680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3695..3834
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3760..3779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3920..3942
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3936..4127)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4936..6018)
                     /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     5980..5999
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6019..6096)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6302..6321
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtcgaaggag aacaagtcgg atgtagtgac
      121 aggtgcctct gaaaccctgg atgcgtccat gcttgaaaat aagaaaacca cgtcgccctg
      181 gccgggcccg cagggtatga tccctgtgac ctcagggaat gcagatgatg cgaatgtgca
      241 caaccgtcac tacctcgatc gtctgttggt tgaaatgcgc gtgatcgatg cagttgagcc
      301 cgatctgacg acgaccattt tcgggaagaa atatgcttcg cctctgatgc cggcagcgtt
      361 gtcacatctt aacaagatct tagacgacaa gaaccgtaaa ccaatgcaag aaaaggccat
      421 tgcggcgcgc gagttgaacc tgctcaactg gatcggcatg gaaacgaacg aggaatactc
      481 agagatcgtt gcggaaggtg gggataccat tcgtattatc aagccgttcg cagatcctca
      541 aaagattatg ggcgaaatta agtttgccga ggaccacggc gcagtggcag tgggcattga
      601 cattgaccat attgcgggcg agaacggtaa gtatgatgta gtagatggta tcccgttagg
      661 gagtatccgc atggatgact taaagaagta cgcagcatca accgaactgc cttttattgc
      721 gaagggcgtt ctgtcggttg cggatgcgtt gaaagcacgt caggccggtt gtaaagcaat
      781 cgtagttagt catcaccacg gccgtgtccc gttcgggatc ccaccgctca gtattctgcc
      841 ggaaatcaag aaggcgctga tcggctccgg aatgcagatc tttgtggacg gctcgttgat
      901 gagtggctat gacgcctaca aagcccttgc cttaggagcg gatgcggttt taattggacg
      961 cggaattctc ccagaggtac tgaagaacgg taccgaggcc acgaagaaca agctccagaa
     1021 aatgaacgag cagttaagcg agatgatgct gtatacgggc atcaaggata tgaacagctt
     1081 cgatcctacg gtgttacact tcgagaaagc ggccgcactc gagcatcatc accatcacca
     1141 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1201 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1261 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     1321 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1381 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1441 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1501 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1561 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1621 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1681 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1741 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1801 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1861 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1921 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1981 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2041 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2101 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2161 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2221 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2281 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2341 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2401 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     2461 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2521 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2581 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2641 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2701 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2761 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2821 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2881 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2941 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3001 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3061 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3121 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3181 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3241 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3301 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3361 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3421 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3481 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3541 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3601 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3661 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     3721 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     3781 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3841 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3901 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3961 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     4021 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     4081 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     4141 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     4201 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     4261 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     4321 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     4381 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4441 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4501 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4561 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     4621 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     4681 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     4741 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     4801 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     4861 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     4921 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4981 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5041 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5101 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5161 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     5221 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5281 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     5341 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5401 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5461 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5521 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5581 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     5641 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     5701 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     5761 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     5821 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     5881 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     5941 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     6001 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     6061 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     6121 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     6181 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6241 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6301 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6361 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaat
//
