LOCUS       sequence-354367-        6626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0W0YYP4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6626)
  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 6626)
  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: A0A0W0YYP4
FEATURES             Location/Qualifiers
     source          1..6626
                     /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             1346..1375
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1428..1446)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1442..1489
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1526..1981
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1613..1632)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1823..1844
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2007..2111
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2112..2180,2181..2972)
                     /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(2330..2349)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3143..3731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3632..3651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3885..3902
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3917..4056
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3982..4001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4142..4164
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4158..4349)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5158..6240)
                     /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     6202..6221
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6241..6318)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6524..6543
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gttaaatcgt cgtaacagct cattatacga
      121 ttacgtgatt atcggagcag gtattgttgg cttaagtgta gctcgtgagc ttttagcgcg
      181 tgaaccgaat gcgaagattg cagttgtgga aaaggaaaat accatcggct tacatgcttc
      241 tggacgcaat agcggtgtgc tgcactcagg gatctactac cagccggata gccttaaggc
      301 aaaggtatgt ctggaaggcg cccgcctcat gagttcgtat tgtgaggaac acggtctgcc
      361 gctgaaccgt attgggaaag tgattgtacc agtccgttgt agcgacggag aaaacctgaa
      421 cgtgctgcac caacgcgcat tatcaaacgg tgcgagtatc cgtttaatta acgatcaaga
      481 attaaaagaa attgaacctc acgcgacaac agccacgggt atggcattac acgcaccgga
      541 aacagccgtg gtggatccta tgttaattat gaagcacctt cactcctcgc tcaccaaacg
      601 cggtgttcac ttcttcttta atagttttat tgatgcgatt gatacacagg ggaagaagat
      661 cacgacaagt gaaggtaata ttagctatgg acatctgttc aatacagccg gcctgttcgc
      721 ggacaagatc gcccacgcgt gtggtctgaa gagcagctat cgcattttac cgtttaaggg
      781 gtactactac gagttgtcac cggacagcaa gctgcaaatc aaccacttaa tctaccccgt
      841 acccgatatg aacgtcccgt tcctgggcat tcatttcacg aagtcagtgt cgggcaccgt
      901 ttatattggt ccgacggcaa tccctgcctt gggccgtgag cattaccgtg gcgtggccgg
      961 catcaaactg tgcgacttca tcaatatcgt gactagtctc ggacgccagt actatgttaa
     1021 tcgccagggt ttccgctcgt acacacatca ggaaatccct cgcatcttta agtctcgttt
     1081 tattaaaagt gcgcgcgcct tagtaccggg cgtcagcaaa tccgacctgc gtgtgtcaaa
     1141 caagatcggt attcgtgccc agttatacga ttataagcat aatgagttga tcaccgactt
     1201 tctgatcagc cgcacggacc acgttactca tgtactgaac gccgtctcac cggctttcac
     1261 tagtgcgttt tcttttgcga agtggattgt taacgagcag aaaacagttt gtaaagaacc
     1321 ggtgtatcgc gcggccgcac tcgagcatca tcaccatcac caccaccacc accactgaga
     1381 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     1441 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     1501 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1561 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1621 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1681 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1741 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1801 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1861 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1921 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     1981 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2041 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2101 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2161 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2221 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2281 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2341 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2401 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2461 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2521 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2581 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2641 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2701 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2761 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2821 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2881 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2941 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3001 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3061 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3121 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3181 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3241 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3301 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3361 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3421 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3481 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3541 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3601 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3661 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3721 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3781 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3841 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3901 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3961 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     4021 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     4081 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4141 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     4201 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     4261 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     4321 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     4381 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     4441 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     4501 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     4561 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     4621 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     4681 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     4741 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     4801 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     4861 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     4921 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     4981 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     5041 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     5101 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     5161 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5221 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     5281 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     5341 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     5401 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     5461 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     5521 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     5581 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     5641 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     5701 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     5761 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     5821 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     5881 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     5941 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     6001 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     6061 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     6121 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     6181 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     6241 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     6301 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     6361 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     6421 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     6481 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     6541 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     6601 ggatcgagat ctcgatcccg cgaaat
//
