LOCUS       sequence-354403-        6581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier S2DJ52.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6581)
  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 6581)
  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: S2DJ52
FEATURES             Location/Qualifiers
     source          1..6581
                     /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             1301..1330
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1383..1401)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1397..1444
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1481..1936
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1568..1587)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1778..1799
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1962..2066
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2067..2135,2136..2927)
                     /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(2285..2304)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3098..3686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3587..3606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3840..3857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3872..4011
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3937..3956)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4097..4119
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4113..4304)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5113..6195)
                     /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     6157..6176
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6196..6273)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6479..6498
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggactttcaa gttatcatca tcggcggcgg
      121 tattgtcggc ttagctacag gtctcaaaat taaacagcgc aatcctaata ttaaggtggc
      181 cctgctcgag aaggaagagg aagtcgcaaa acaccagacg ggtaacaact cgggagtgat
      241 ccactcgggc ctgtactaca agccgggcag cttaaaagcg aagaattgca ttgaaggcta
      301 tcatgaactc gtacgctttt gtgaggaaga gaatattccc tttgagctga ccggcaaggt
      361 cgtggtggcg acccgcaagg aacaagtgcc cttgcttaac tcgctgcttg aacgtggtct
      421 gcagaacggt ctcaaaggca cccgcagtat cactctcgat gaattaaagc atttcgagcc
      481 ttattgcgcc ggcgtagcag ctatccatgt gccgcagacc gggattgttg actataagct
      541 cgtggcggaa aagtacgccg agaaatttca aatcctgggc ggccaggtgt ttctgggcca
      601 caaggtcatt aaggtcgaga cccagaacac tgcgtctatt attcatacca gtaaggggag
      661 cttttcaacc aacttgctga tcaattgcgc aggcttatac tctgacaaag ttgcgcaaat
      721 gaaccagaaa gagagtctgg acgtcaagat cattccgttt cgcggcgagt actataagat
      781 taagaaagaa cgtgagtact tagtgaagaa tctgatctac cctgtgccgg atcctaattt
      841 tccatttctg ggcgtgcatt tcacccgcat gatgaaaggc ggcgtagaag ccggcccaaa
      901 cgctgttctc gccttcaaac gtgaaggata taagaagtcc caagtaaatt tctctgaatt
      961 ggccgaaacg ctgtcgtggc caggattcca aaaggttgcg tccaaatatt ggaaaacggg
     1021 catgggtgag ctgttccgct cgttttctaa gaaagccttc acggatgcct taaaagaact
     1081 catcccggat attcaggaaa gcgatttgat cgaaggcggt gccggcgtgc gcgcacaagc
     1141 atgtgaccgc accggtgggc tgttggatga tttctgcatt cgcgaagatc agaatgctat
     1201 tcatgttctc aacgccccgt cgccagcggc gacctcttcg ttgagcatcg gcggcactgt
     1261 atgcgaatgg gcgcttaaac gtttcgcggc cgcactcgag catcatcacc atcaccacca
     1321 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1381 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1441 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1501 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1561 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1621 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1681 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1741 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1801 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1861 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1921 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1981 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2041 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2101 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2161 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2221 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2281 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2341 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2401 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2461 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2521 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2581 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2641 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2701 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2761 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2821 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2881 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2941 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3001 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3061 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3121 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3181 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3241 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3301 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3361 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3421 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3481 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3541 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3601 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3661 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3721 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3781 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3841 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3901 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     3961 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4021 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4081 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4141 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4201 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4261 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4321 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4381 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4441 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4501 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4561 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4621 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4681 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4741 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     4801 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     4861 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     4921 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     4981 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5041 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     5101 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5161 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     5221 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     5281 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     5341 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     5401 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     5461 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     5521 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     5581 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     5641 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     5701 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     5761 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     5821 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     5881 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     5941 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     6001 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     6061 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     6121 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     6181 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     6241 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     6301 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6361 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6421 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6481 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6541 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa t
//
