LOCUS       sequence-354438-        6980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0P7WDX3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6980)
  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 6980)
  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: A0A0P7WDX3
FEATURES             Location/Qualifiers
     source          1..6980
                     /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             1700..1729
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1782..1800)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1796..1843
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1880..2335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1967..1986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2177..2198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2361..2465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2466..2534,2535..3326)
                     /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(2684..2703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3497..4085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3986..4005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4239..4256
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4271..4410
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4336..4355)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4496..4518
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4512..4703)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5512..6594)
                     /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     6556..6575
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6595..6672)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6878..6897
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaacgttcca gccaccccat cccctgatgg
      121 aaccgcagag gccgcccctg cactggttgc aggtctcgaa gctttgctgg gtgcagaagc
      181 agtcgaaacc gatgaagaag agaggcgatt cgctgttcag gacttgttct tcaaaggggc
      241 tttaccatta gccgtagtat caccagggtc tacaagagaa gtgagcgaat tggttcgttg
      301 gtgtgctaag caagccgtgg ccatagctcc ccgtggcggc gggatgagct acacagacgc
      361 ctttcagcct agagttcaca gatcggttgt tttgggattc tctcgattag ctgccataag
      421 atcaatcgac ccagccgacg gaaacgtaac ggttgaggct gggtgtacat gggcagctct
      481 tgacgcagca cttgctaaag tcggtatgag ggctcgtttc tggggtccta tgtcaggagc
      541 tgccgctact attggtggtt ctttaagcca gggttccgtg actttcgggt caggtgttgc
      601 tggtgcttct gctaacgcag ttaaatcttt cgaaatcgtt acgggtacgg gtgaagtttt
      661 acatacggga agcgacggct ctgctgggac aggtcccttt aatcgaaact ttggtccaga
      721 ctttaccggt ttgtttgcta acgatgctgg ggctctgggc attaaaactg cagctactat
      781 cgagattgag cccagaccag agcgtgtgga cgggatatca ttcgctgtag attccttcag
      841 ggtcatggcc gaccttttgc gtgatgtgag attgcgacgt ctggcttctg agataatcgc
      901 aatggatgct gaagtcgcaa gacaaaacgc aggtccgtcg tcgctttccg aagacgccaa
      961 ggccctgata agaatcgcta gatctgctgg ttcggcccct gcaggattag caaggatggc
     1021 ccgaattgct gctgccggta gacgatttct agatggtgcc cgatatacag tccactttgt
     1081 agttgaggga agagacagaa gagagttacg ttcacgactc gctgctatcc gaggattggc
     1141 caaaggggca cgtgagatag ttaatactgt tcctctggcc gtcagggctg ctccattccc
     1201 tgaactgcct gtcactcatc cagatggaag aagattgctt ccaattcatg gtgttttcgc
     1261 cgactccagg attgccgcat ttcatacaga ttatctggct ttaaaagctg agcacgctgc
     1321 caggatggca caggctggag taactctcgc tgaattcttc gcaggtgtcg caggtgtcgg
     1381 aaccctatac gaaccggttt tctactggcc agactctgct atgctgtacc atagaaggag
     1441 aactcctgcc tatcttgatg gagtggctaa ggaccacccg gataatcctg ccgctcgagc
     1501 tttggttcga gagatggtcg gagctatcat agctttaatg cgagcccatg gtgccaccca
     1561 tttccaaata gggagactat acccttatgc cgaaaccaga gagggagaag ctggtcgttt
     1621 gctcagagac ctgaaacagc gtctcgaccc tcacggaatc ctgaatccgg gcacccttgg
     1681 gttagctgct gctttagaac accaccatca ccaccaccac caccaccact gagatccggc
     1741 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1801 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     1861 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1921 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1981 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2041 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2101 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2161 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2221 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2281 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     2341 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2401 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2461 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2521 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2581 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2641 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2701 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2761 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2821 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2881 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2941 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3001 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3061 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3121 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3181 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3241 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3301 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3361 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3421 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3481 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3541 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3601 cgaaggtaac tggattcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3661 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3721 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3781 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3841 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3901 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3961 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4021 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4081 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4141 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4201 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4261 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4321 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     4381 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4441 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4501 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     4561 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4621 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4681 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4741 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4801 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4861 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     4921 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     4981 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     5041 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     5101 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     5161 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     5221 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     5281 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     5341 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     5401 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     5461 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     5521 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5581 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     5641 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     5701 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     5761 gtcttcggta tcatcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     5821 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     5881 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     5941 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     6001 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     6061 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     6121 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     6181 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     6241 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     6301 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     6361 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     6421 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     6481 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     6541 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     6601 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     6661 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     6721 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     6781 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     6841 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     6901 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     6961 agatctcgat cccgcgaaat
//
