LOCUS       sequence-354470-        6629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier E6SCX5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6629)
  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 6629)
  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: E6SCX5
FEATURES             Location/Qualifiers
     source          1..6629
                     /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             1349..1378
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1431..1449)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1445..1492
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1529..1984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1616..1635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1826..1847
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2010..2114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2115..2183,2184..2975)
                     /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(2333..2352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3146..3734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3635..3654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3888..3905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3920..4059
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3985..4004)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4145..4167
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4161..4352)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5161..6243)
                     /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     6205..6224
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6244..6321)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6527..6546
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtcaacccgc cgccgtcgtc cgcgctggtc
      121 cgaactgcgt ccactcttac gcattgaacc acctcatctg ggcagcaccc gtcgtcgttt
      181 ggaagcggca gctagcatcg acgatctgcg ccgtattgct cgccgccgta cacctcgtag
      241 cgtgtttgat tatgttgacg gcgcagcaga gcaagaaatt tctttacgtc gtgctcgcga
      301 agcttttagt cgtattgaat ttcgtccacg tgtgttgcgc gatgtgagcg aagttgatgc
      361 gagccgcgtg gtagttggaa gcccgtcatc actgccgttg gttcttgccc ctaccgggtt
      421 cacgcgtatg atgcatcatg aaggtgagcg tgcggttgct cgcgcggcgg cccgtgctca
      481 aattccgtat gcgcttagta ccatggggac cgtttctgta gaagaggttg cggctgcggc
      541 accggggtcc gaactttggt ttcaactcta tctgtggaaa gatcgtgccg cgagtctgga
      601 attagtgcaa cgtgccgcgg cggcaggtta ccgcacttta gtgcttacag tagatactgc
      661 ggttgcggga cgtcgtttgc gcgatgtacg taatggctta acgattcccc ctgctctgac
      721 cgtgcgcact cttgcggaca tgtctttgca tccggcgtgg tggattaact tactgaccac
      781 tgaaccgtta gaattcgcat cactgcgtga ttcaggtggg actgtagctg atctggttga
      841 tcgtatgttt gatcccagcg catccatttc tgatttagcg tggattcgtg atcaatggcc
      901 aggtcgtatt gttgttaaag gtatccaaca tccggatgat gcagtcgcca tggtggatct
      961 gggcattgat gggattattg tttctaacca tggtggtcgc caactggatc gcgcggcgac
     1021 cccgttagaa gttcttcccg atatcgtggc tgcggttgcg ggacgtattg aggttctgct
     1081 ggacactggc attactgatg gtgccgatat tgtagcggcc gtagcaaatg gcgccacggg
     1141 ttgtttagta ggtcgtgcat acttgtatgg tctgatggcc ggcggtgaag caggtgtaga
     1201 tcgtacgctt tcgattctgc gcgatcaagt gactcgtacg atgcgtctct tgggtgtgag
     1261 cagtcttgat gaattgacgc cagagcacgc ggtaattgcg ggttcgggtc ctcgcgcacg
     1321 cgctcaaaca ccggcggccg cactcgagca tcatcaccat caccaccacc accaccactg
     1381 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1441 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1501 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     1561 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1621 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1681 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1741 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1801 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1861 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1921 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1981 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2041 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2101 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2161 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2221 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2281 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2341 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2401 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2461 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2521 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2581 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2641 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2701 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2761 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2821 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2881 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2941 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3001 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3061 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3361 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3421 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3481 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3541 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3601 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3661 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3721 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3781 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3841 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3901 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3961 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4021 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4081 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4141 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4201 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     4261 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4321 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     4381 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     4441 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     4501 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     4561 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     4621 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     4681 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     4741 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4801 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     4861 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     4921 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     4981 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     5041 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     5101 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     5161 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5221 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     5281 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     5341 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     5401 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     5461 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     5521 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     5581 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     5641 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     5701 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     5761 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     5821 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     5881 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     5941 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     6001 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     6061 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     6121 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     6181 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     6241 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     6301 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     6361 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     6421 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     6481 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     6541 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     6601 agaggatcga gatctcgatc ccgcgaaat
//
