LOCUS       sequence-354436-        6809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier D4MUV9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6809)
  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 6809)
  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: D4MUV9
FEATURES             Location/Qualifiers
     source          1..6809
                     /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             1529..1558
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1611..1629)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1625..1672
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1709..2164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1796..1815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2006..2027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2190..2294
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2295..2363,2364..3155)
                     /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(2513..2532)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3326..3914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3815..3834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4068..4085
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4100..4239
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4165..4184)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4325..4347
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4341..4532)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5341..6423)
                     /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     6385..6404
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6424..6501)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6707..6726
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtctgaatac cagtacaata aggtgacgcc
      121 tgagatgatt gagaaattca aggaaatcgc tcctaaaagg gtattagttg gagacgaaat
      181 caatgaagac tttacccacg atgaaatggc catctatgga aaggctagac ctgaagtcct
      241 cgttgaggct acttcaacag aggaagtcgc agcagttgta aagctatgta atgaaaacaa
      301 gatccctgtt accccatccg gtgcacgaac aggcttggta ggtggtgctg tttccatagg
      361 tggtggagtt atgatatcac tgacaaagat gaacaaaatc ttgggttatg acaaggagaa
      421 tttcgttgtt aagatacagt ctggagtttt gctgaatgat ctcgcacagg atgcagagaa
      481 gcaaggtctt ttgtacccac ctgacccagg agagaaattt gctacagttg gtggtaatgt
      541 cgccacaaac gccggtggta tgcgagccgt caaatatggt tgtacacgag actacgttag
      601 ggccatgaca gttgtacttc ctaccggaga gatcgtcaag ttgggagcca ctgtctcaaa
      661 gacttcctcg ggttactcat tgcttaatct gatgatcgga tcagagggaa ctttgggaat
      721 tatcacagaa cttactttaa aggttattcc tgccccaaaa tccgtgataa gcttgattat
      781 cccatacgaa aacctcgagg actgtatcgc aactgtccct cagttcttca tgcatcattt
      841 ggctccgcaa gccttggagt ttatggagaa agaggttgtt atggatactg aaaagttctt
      901 agggaagcaa gtttatccga aggagctcga aggtactgag attggtgctt atctattagc
      961 tacatttgat ggtaattctg aggaacaatt agaggacatc atagaacagg cttccgaggt
     1021 ggtactagag gcaggtgcca ttgacgtctt ggttgcagat acaccagctt taaagaaaga
     1081 tgcctgggcc gtcagaggag ccttactcga agccattgaa gctgatacag tcttattaga
     1141 cgagtgcgac gttgtcgttc caactaacaa gatcgctgaa ttcctgacat acactaagtc
     1201 attggaagct gaggctgact tcagagtaaa gtcctttggt cacgctggcg atggaaacct
     1261 acacatttat acctgttcga atgacatgga agagggtgag tttaagaagc aagtggctgt
     1321 gtttatggat aaggtgtacg ctaaggccac agagttcggt ggcatgatca gtggagaaca
     1381 tggaatcggt cacggcaaaa tggattactt ggccgaatcc ctgggtcctg ttcagatgag
     1441 aatcatggaa ggcgttaaag aggttttcga tcctaacatg atcctgaacc cgggtaagat
     1501 atgctacaaa ctcgctgcag ctctggaaca ccaccatcac caccaccacc accaccactg
     1561 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1621 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1681 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     1741 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1801 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1861 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1921 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1981 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2041 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2101 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     2161 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2221 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2281 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2341 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2401 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2461 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2521 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2581 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2641 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2701 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2761 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2821 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2881 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2941 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3001 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3061 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3121 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3181 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3241 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3301 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3361 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3421 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3481 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgttc
     3541 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3601 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3661 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3721 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3781 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3841 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3901 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3961 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4021 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4081 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4141 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4201 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4261 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4321 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4381 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     4441 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4501 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     4561 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     4621 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     4681 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     4741 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     4801 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     4861 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     4921 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4981 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     5041 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     5101 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     5161 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     5221 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     5281 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     5341 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5401 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     5461 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     5521 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     5581 acatgagctg tcttcggtat catcgtatcc cactaccgag atatccgcac caacgcgcag
     5641 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     5701 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     5761 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     5821 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     5881 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     5941 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     6001 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     6061 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     6121 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     6181 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     6241 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     6301 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     6361 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     6421 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     6481 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     6541 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     6601 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     6661 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     6721 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     6781 agaggatcga gatctcgatc ccgcgaaat
//
