LOCUS       sequence-354466-        6110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier I9UMF5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6110)
  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 6110)
  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: I9UMF5
FEATURES             Location/Qualifiers
     source          1..6110
                     /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             830..859
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(912..930)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      926..973
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1010..1465
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1097..1116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1307..1328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1491..1595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1596..1664,1665..2456)
                     /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(1814..1833)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2627..3215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3116..3135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3369..3386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3401..3540
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3466..3485)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3626..3648
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3642..3833)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4642..5724)
                     /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     5686..5705
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5725..5802)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6008..6027
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaaggtaaac ttcttcgcca cgtgccttgg
      121 tgctgctata tactccaacg caagcctcaa cgctattaag ctcctgagga aggaaaactt
      181 agaagtagta ttcaagaaag accagacatg ctgttcgcag ccatcataca acagcggata
      241 ctatgaagag actaagaaag tagtgttgta taacattaaa ctgtactcca ataatgatta
      301 tcccatcatc cttccatctg ggtcttgtac cgggatgatg aggcacgatt acctggaatt
      361 gtttgaaggt cacgccgaat tcaatatggt taaggacttc tgttctcgag tgtacgaatt
      421 gagtgaattc ctagacaaga aactgcaagt gaagtacgag gacaagggtg aacctttgaa
      481 gatcacctgg cactcaaact gtcacgctct gagggtggct aaagtgatcg atagtgccaa
      541 gaaccttatt agacaactga agaacgtgga attgatagag ttggagaaag aagaggaatg
      601 ctgcggtttc ggtggaactt tctcagtcaa agaacccgag attagcgcag ttatggttaa
      661 agagaagatc aaggatatag agagccgtca tgtcgatgtg atcgtcagcg ctgacgccgg
      721 atgcttgatg aacatcagca cagctatgca gaaaatgggc agcctaacca agcctatgca
      781 tttctatgac ttcctcgcat ccagacttgg tttggcagca gccttagaac accaccatca
      841 ccaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
      901 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
      961 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     1021 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1081 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1141 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1201 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1261 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1321 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1381 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     1441 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1501 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1561 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1621 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1681 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1741 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1801 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1861 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1921 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1981 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2041 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2101 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     2161 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2221 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2281 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2341 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2401 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2461 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2521 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2761 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     3421 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     3481 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     3541 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     3601 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     3661 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     3721 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     3781 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     3841 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     3901 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     3961 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     4021 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     4081 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     4141 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     4201 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     4261 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     4321 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     4381 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     4441 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     4501 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     4561 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     4621 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     4741 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     4801 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     4861 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     4921 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     4981 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     5041 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     5101 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     5161 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     5221 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     5281 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     5341 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     5401 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     5461 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     5521 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     5581 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     5641 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     5701 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     5761 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     5821 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     5881 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     5941 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     6001 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     6061 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
