LOCUS       sequence-345535-        6875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A9D099.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6875)
  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 6875)
  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: A9D099
FEATURES             Location/Qualifiers
     source          1..6875
                     /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             1595..1624
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1677..1695)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1691..1738
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1775..2230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1862..1881)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2072..2093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2256..2360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2361..2429,2430..3221)
                     /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(2579..2598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3392..3980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3881..3900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4134..4151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4166..4305
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4231..4250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4391..4413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4407..4598)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5407..6489)
                     /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     6451..6470
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6490..6567)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6773..6792
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gactgatctc atcgctttct tggaaccttc
      121 ccgatccgtt atcgatagaa gaccaactat catagcagac cttgcagaat tattgcctga
      181 gggatgtctg atcactgaag aacgagagct ggtcccattc gacaccgatg cttttgtctc
      241 atatagacgt atgcctttag ctgttgtgtt acctgaaact accgcccagg tggccgctgt
      301 gatgaaatac tgtaacgaca acgcaattcc tgtaataccg cgtggagcag gaacgagctt
      361 gtcaggcgga gctatcccac aagaggacgc tgtggtaatt ggcgtcagca agatgaatca
      421 gatagttgag gtggactacc ctaatcgaac agctgttgtg caggccggtg ttaccaacat
      481 aagcatctct caaagtgttg atgcagacgg cttcttctat gcacctgatc cttcttctca
      541 gcttgcctgt actattggtg gtaatatcgg aatgaactcc ggtggtgctc actgcctgaa
      601 atatggtgtt accactaata atctccttgg agtcaaaatg gtcatgtttg acgggtccat
      661 tgtcgaaatc ggtggaaagg ccctagattc cgcaggctac gatcttctgg gacttatctg
      721 tggcagtgaa ggacaacttg gtatcgtgac agaagccact gtgagattgc tggccaagcc
      781 agaaggtgct aggccagtcc tcttcggttt cgagagttca gtggctgctg gggcttgtgt
      841 aaccgaaatc attggatctg gaataattcc cgttgctatt gaatttatgg ataagcctgc
      901 tattgagata tgtgaagcat ttgctcaagc cggatatcca atggatgttg aggctttact
      961 aatcgtggaa gtagagggat cagacgctga aatggatgcc atgttggcta gaataatcga
     1021 aatagcccga aagcacgccg tgaagactat caaggagtcc caatccgcca ctgaggctgc
     1081 cctgatctgg aaaggaagaa agtctgcctt tggtgctaca ggaaggattg ctgactacat
     1141 ctgtatggat ggcaccgttc ctttaggaca gttgtcccat gtcctagaaa agacatccga
     1201 aatcatagac cactacggtt taagagtagc caacgttttc cacgctggcg acggtaacat
     1261 gcaccccctg atcctatttg acgcaaatga cccagaggaa tcaatgaaag ccgaaatggc
     1321 tggaaatgac atacttaaat tgtgtgtaga tgcaggtggt tgtttgactg gagagcacgg
     1381 agtaggtatt gagaagaggg acttgatgcg tcaccaatac actgatgctg acttggctca
     1441 gcagatgaga gttagggctg cctttgatcc gcagtggatc ctcaatcctt ctaaagtgtt
     1501 tccattggag agacgtaacg gatcgcaacc tggtgatcca gttcctgacg cagcagcagc
     1561 tcctgggaaa actgcagctg ccgctgctct ggaacaccac catcaccacc accaccacca
     1621 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1681 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1741 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1801 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1861 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1921 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1981 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2041 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2101 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2161 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2221 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2281 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2341 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2461 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2521 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2581 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2641 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2761 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2881 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     2941 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3181 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3241 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3301 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3361 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3661 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3721 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3781 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4021 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4081 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4141 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     4201 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4261 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     4321 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4381 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4441 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     4501 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     4561 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     4621 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     4681 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     4741 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     4801 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     4861 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     4921 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     4981 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     5041 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
     5101 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     5161 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     5221 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     5281 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     5341 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     5401 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5461 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     5521 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     5581 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     5641 gatataacat gagctgtctt cggtatcatc gtatcccact accgagatat ccgcaccaac
     5701 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5761 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5821 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     5881 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     5941 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     6001 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     6061 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     6121 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     6181 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     6241 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     6301 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     6361 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     6421 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     6481 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     6541 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     6601 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     6661 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6721 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6781 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     6841 cggcgtagag gatcgagatc tcgatcccgc gaaat
//
