LOCUS       sequence-354325-        6545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0T9WB25.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6545)
  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 6545)
  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: A0A0T9WB25
FEATURES             Location/Qualifiers
     source          1..6545
                     /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             1265..1294
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1347..1365)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1361..1408
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1445..1900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1532..1551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1742..1763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1926..2030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2031..2099,2100..2891)
                     /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(2249..2268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3062..3650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3551..3570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3804..3821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3836..3975
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3901..3920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4061..4083
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4077..4268)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5077..6159)
                     /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     6121..6140
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6160..6237)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6443..6462
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaccgagaag aataatatta attatgacgt
      121 gatcattatc ggtagtggta tcattggcac caacatcgcg tacgaactta gcaaatttga
      181 actgaaaatc gccttacttg aggcaaagaa ataccccgcg agcgagacca ctaccggcaa
      241 ctcaggtatt atccattgcg gtttcgatcc tagtccgggt aagctgaagg ccaaattcaa
      301 cgtcttaggt cacaagttgt ggaccgagaa catcttcaag aaaatcattt tcccgcgcca
      361 gaaagcgaag agtgtagttc tggcctttga ttccgaggaa gagaaaatta tctacgatct
      421 ttacaagcgt ggcatcatta acggtctcga tccgaagaat ttgagcatcc tgagtaagaa
      481 ggagctgaag ctggcccatc cggaagtgag tgaggaagcg gtgttagccc tcgaatgcct
      541 gaactcgtgg gtcgtggacc ctgtcaaggc cgcatatgca tttcttgaaa ttagtcgcaa
      601 gaacggcctg gaattcttct cgtcctctaa ggtcatctcc atttcgaaga actcgcaaaa
      661 tttcttctta attaagacgg aaaaccagaa taaaccgttc ttccaggcga agattattat
      721 taacgcggcg ggccattacg cggacgaaat cgccaaaatt gcggggtttg gcgaattctc
      781 tcagaagcag cgccgcggac aatatctgat tctcaagaac catccaaata ttaagaatat
      841 ttattttctg gtcccttcta aatatggaaa aggtgtaatc gtcgccccgc gtcctgacgg
      901 caatatcctg gtcggcccga ccgcgaaaga gaacatccaa aagaacaagg cgaattgtct
      961 ggactttaac cttaacgaag aaattcagaa aattggccat aaaattatcc cgagtctgga
     1021 ttacagtgaa acggtatttg cactcgctgg ttctcgccca atcgaagcgg taaagagcga
     1081 ctttgtgatt aagcccgcgt ttaatgatcc taattttatc ttcgcggcgg ggatgcagtc
     1141 cccgggtctg tcagcctctc cggctatcgc gttagagatt gttaattttg tcaagaaccg
     1201 tatcgactta gtgaagaaga ccgagctggt tgaaacgatt gaatatattg cggccgcact
     1261 cgagcatcat caccatcacc accaccacca ccactgagat ccggctgcta acaaagcccg
     1321 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1381 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     1441 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1501 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1561 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1621 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1681 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1741 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1801 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1861 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     1921 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1981 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2041 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2101 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2161 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2221 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2281 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2341 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2401 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2461 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2521 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     2581 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2641 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2701 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2761 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2821 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2881 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2941 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3001 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3061 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3121 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3181 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3241 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3301 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3361 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3421 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3481 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3541 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3601 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3661 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3721 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3781 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3841 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3901 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3961 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4021 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4081 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     4141 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     4201 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4261 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4321 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4381 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     4441 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     4501 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     4561 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4621 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4681 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     4741 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4801 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4861 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4921 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     4981 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     5041 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5101 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5161 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5221 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5281 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5341 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5401 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5461 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5521 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5581 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5641 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5701 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5761 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5821 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5881 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     5941 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6001 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6061 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     6121 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6181 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6241 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6301 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     6361 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6421 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6481 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     6541 gaaat
//
