LOCUS       sequence-354357-        6005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A090SY56.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6005)
  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 6005)
  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: A0A090SY56
FEATURES             Location/Qualifiers
     source          1..6005
                     /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             725..754
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(807..825)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      821..868
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      905..1360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(992..1011)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1202..1223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1386..1490
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1491..1559,1560..2351)
                     /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(1709..1728)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2522..3110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3011..3030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3264..3281
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3296..3435
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3361..3380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3521..3543
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3537..3728)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4537..5619)
                     /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     5581..5600
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5620..5697)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5903..5922
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggcgaaggtg catgacttta ttgtgattgg
      121 cggtggaatt gttggagtga gcacggcatg gcaacttcaa cgcaagtacc cccacgcttc
      181 aatcgtgctc gtcgagaaag agagtaattt tgcaaagcat cagacgggtc acaactcagg
      241 cgtaattcat gctggcgtgt attatcagcc tggctcactg aaagcagact tttgcaagcg
      301 cggtgctgaa gccactaaac gtttctgtgc gcaacataac atcaaagtgg aaaattgcgg
      361 gaaactgctt gtagccacca atgccgtgga gaagcagcgc atggatgccc tgtacgaacg
      421 ctgcatcgaa aacgggattg acgttagtct tttgagtcaa acggaattag aggagattga
      481 gccaaatatc actggcctgg gcgccattaa agtagaggcg acctcgattg tggattaccg
      541 cctggtcaca cagaccatgg cggaacagtt ccagcttttg ggtggagaag ttttactcag
      601 cactgaagtc accgcgctga cggaacagca ggattggtct atcagcattg caagtccaat
      661 ggagcgtcaa cctttatgta aactggcgaa tctctcatgc gcggctggag cggccgcact
      721 cgagcatcat caccatcacc accaccacca ccactgagat ccggctgcta acaaagcccg
      781 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
      841 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
      901 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
      961 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1021 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1081 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1141 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1201 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1261 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1321 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     1381 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1441 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1501 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1561 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1621 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1681 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1741 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1801 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1861 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1921 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1981 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     2041 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2101 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2161 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2221 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2281 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2341 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2401 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2461 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2521 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2581 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2641 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2701 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2761 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2821 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2881 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2941 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3001 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3061 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3121 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3181 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3241 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3301 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3361 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3421 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3481 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3541 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3601 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     3661 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     3721 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     3781 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     3841 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     3901 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     3961 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     4021 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4081 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4141 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     4201 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4261 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4321 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4381 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     4441 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     4501 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4561 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4621 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     4681 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     4741 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     4801 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     4861 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     4921 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     4981 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5041 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5101 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5161 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5221 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5281 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5341 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     5401 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     5461 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     5521 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     5581 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     5641 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     5701 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     5761 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     5821 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     5881 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     5941 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     6001 gaaat
//
