LOCUS       sequence-354400-        6029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier B7PPE8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6029)
  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 6029)
  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: B7PPE8
FEATURES             Location/Qualifiers
     source          1..6029
                     /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             749..778
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(831..849)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      845..892
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      929..1384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1016..1035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1226..1247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1410..1514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1515..1583,1584..2375)
                     /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(1733..1752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2546..3134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3035..3054
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3288..3305
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3320..3459
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3385..3404)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3545..3567
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3561..3752)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4561..5643)
                     /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     5605..5624
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5644..5721)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5927..5946
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gatcctcgga gttgaatcag gtgtgcttcg
      121 ccgcttacgt ttacgcccac gtatgttacg tggcgtcaaa gaccgcgata tgacggtgac
      181 ggtgctgggc cgccaacgcc tgagcatgcc attaggcatc gctccgtccg cgatgcaacg
      241 tatggctcat cccgacggtg aagaggcaac cgcaaaagct gctgagaaag ccggaacagt
      301 tatgattctg tcaactctgt ctacgactag catggaagac gtgcgcaaag ccgccccgca
      361 cgccatcttg tggtatcagc tctacgtgtt tcaagatcgt gaactcaccc gtcgcttggt
      421 taaacgcgcg gaacaggcgg gatattcagc gttagtgttg acggttgatg cgccggtttt
      481 cgggcgtcgc gtcagtgacg tccgtaaacg tttctcgtta ccctcgcatt taaagtatgt
      541 gatctgctgt cgccagagcc cattcctttt cttctttgta ttcttttgtt tcatgggttg
      601 ctttgaaaag atgcatctga ttaatttcat ctccgcggaa gacgcggttt tagctgtacg
      661 taacggcatc cccgccatca tcgtcagcaa ccacggcggc cgtcagctgg acggtgtcgc
      721 ctcgacggtg tctgcggccg cactcgagca tcatcaccat caccaccacc accaccactg
      781 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
      841 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
      901 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
      961 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1021 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1081 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1141 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1201 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1261 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1321 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1381 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1441 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1501 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1561 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1621 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1681 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1741 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1801 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1861 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1921 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1981 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2041 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2101 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2161 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2221 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2281 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2341 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2401 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2461 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2521 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2581 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2641 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2701 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2761 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2821 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2881 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2941 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3001 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3061 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3121 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3181 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3241 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3301 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3361 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3421 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3481 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3541 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3601 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3661 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3721 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3781 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     3841 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     3901 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     3961 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     4021 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     4081 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     4141 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4201 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     4261 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     4321 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     4381 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     4441 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     4501 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     4561 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4621 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     4681 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     4741 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     4801 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     4861 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     4921 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     4981 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     5041 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     5101 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     5161 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     5221 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     5281 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     5341 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     5401 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     5461 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     5521 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     5581 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     5641 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     5701 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     5761 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     5821 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     5881 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     5941 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     6001 agaggatcga gatctcgatc ccgcgaaat
//
