LOCUS       sequence-354430-        6359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier Q96Y72.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6359)
  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 6359)
  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: Q96Y72
FEATURES             Location/Qualifiers
     source          1..6359
                     /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             1079..1108
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1161..1179)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1175..1222
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1259..1714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1346..1365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1556..1577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1740..1844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1845..1913,1914..2705)
                     /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(2063..2082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2876..3464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3365..3384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3618..3635
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3650..3789
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3715..3734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3875..3897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3891..4082)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4891..5973)
                     /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     5935..5954
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5974..6051)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6257..6276
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gggtcttgag gaagagttaa agaagtgtgt
      121 ccattgcggg ttttgccttg aggcctgccc cacctacgtt gtcactcgtt ctgaggtaca
      181 tagtcctaga ggacgtatta ctgcagtgaa gttaaacata ctctctgaag gtataaatac
      241 ttgcatgtac tgcagaaggt gcgagctggc ctgcccatct ggtgtgatct attctgaaat
      301 tatcaccaga gttaggaaac ccaatacaat ccaaaaggtt atgttgcgat tattggagcg
      361 tcctcatata ctcccatact ttatccgtgg tgagtatgct aagaaaattc cggtgaaggc
      421 cgagaagcct atagaatata gagacaacaa cgaagaaatc atcttgttcc ccggttgtat
      481 aactagcata ttctttaaag agacggtcca gaagacgttg aattatttga agagcttggg
      541 gtaccgtgtg aagatctaca atggttgttg tggtttagcc cacaagaacg ccggggaact
      601 taacagatcg aaagagctca tcgagaagct gagaaaggaa ttccacggta gaacaatcgt
      661 gtcgctgtcc tcgaactgct cagctcacat gaaagaaaat ggtctggaag tttacgattt
      721 ccccgaattc atcataaagt ttaaccttcc cttgccgaag gtagagagac gtcttaccat
      781 ccatgaacca tgccacgcta atattttggg cctaaacaaa tatactcgtg aggtgttgtc
      841 taagatgggt gtagagatca gtgagagcga ggagccttcg ttcgaatgcg gtgcaggtgg
      901 agactatttc ttatttcatg aggaaatctc tgagaaggta atgcaggtga agaaggaaaa
      961 gacgttgaag tctggtgtta acacagtagt ctcgacaaac ccatcctgct ccctagcctt
     1021 catcaagatg ggcttgaagc caatacacat cgctgactta ttggccgctg ctctggaaca
     1081 ccaccatcac caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     1141 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1201 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     1261 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1321 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1381 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1441 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1501 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1561 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1621 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1681 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1741 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1801 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1861 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1921 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1981 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2041 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2101 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2161 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2221 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2281 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2341 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     2401 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2461 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2521 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2581 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2641 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2701 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2761 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2821 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2881 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2941 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3001 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3061 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3121 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3181 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3241 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3301 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3361 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3421 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3481 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3541 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3601 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     3661 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     3721 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     3781 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3841 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3901 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     3961 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4021 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4081 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4141 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4201 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4261 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4321 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4381 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4441 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     4501 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     4561 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     4621 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     4681 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     4741 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     4801 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     4861 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4921 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4981 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5041 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5101 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat catcgtatcc
     5161 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5221 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5281 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5341 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     5401 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     5461 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     5521 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     5581 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     5641 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     5701 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     5761 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     5821 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     5881 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     5941 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6001 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6061 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     6121 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6181 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6241 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     6301 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaat
//
