LOCUS       sequence-354394-        6569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A021XIZ9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6569)
  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 6569)
  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: A0A021XIZ9
FEATURES             Location/Qualifiers
     source          1..6569
                     /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             1289..1318
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1371..1389)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1385..1432
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1469..1924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1556..1575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1766..1787
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1950..2054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2055..2123,2124..2915)
                     /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(2273..2292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3086..3674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3575..3594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3828..3845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3860..3999
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3925..3944)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4085..4107
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4101..4292)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5101..6183)
                     /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     6145..6164
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6184..6261)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6467..6486
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gattttcgat tattgcattg taggaggagg
      121 tattgtgggc ctcgcgaccg caatgcgtct gttggagaat aaaccgggtt cctcattgat
      181 cttgatcgag aaagaggacg cgcttggcaa gcatcagacg ggccataata gtggtgttat
      241 ccacgccggc atttactacc cacctggttc tttgaaagcg gagttatgtc gcaagggcgc
      301 agaggcgacc aaagcgttct gctctgagaa cggtatccgc tttgaagtgt gtggcaaact
      361 tctcgtcgcc acttctgacc tggagattca gcgcatggac gcgctgtacg agcgtagcaa
      421 gcagaatacg atcgaagtac atcgtgtgtc tgagggagag ctccgcgagc gcgaaccaca
      481 cattcgtggg cgtggagccc tgtttgtgcc gagcaccggc atcgtatcgt acgcggaagt
      541 ttgccaagcg atgggtcgcc gcattaaggc gctgggtggt aatatccgct tatctacccg
      601 cattacaggc atccgtgaaa cgattgacgt agttgacatc accagcgatg gccaaacttg
      661 gcaagcaaag aaacttgttg cgtgtgcagg ccttcaatct gaccgtctcg cagtactggc
      721 cggactgagc attgagcatc gtatcgtccc ttttcgcggg gaatattttc gtttgcctgc
      781 ggacaagaac gacatcgttc gtcatcttat ctaccctatt ccggatccgg acttgccctt
      841 tctgggagta cacctcaccc gtatgattga tggttcggtt accgttggcc cgaatgcagt
      901 gatcggcttc gcgcgcgagg gctacccacg cctgtcaatc aatcttgccg atatggccga
      961 ttacgccctt ttcccgggtt tctggaaaac cgtcttcgcc aaccgtaagt ctgccgtcac
     1021 cgagttgctg aacagcctgt ggaagcccgg ctatctggcc gaatgcaaga agtattgccc
     1081 cagtttgacg ctcagtgatc tgctgccata cgaggcaggt attcgcgcgc aagccgttcg
     1141 taaagatggc actctgatcc acgactttct gttcgcagag accgatcgta tgctgcatgt
     1201 gtgcaatgca ccgtcgccgg cggctaccag cgcgattccg attgcggaga tgattgttgg
     1261 gcgcctgggc aacgcggccg cactcgagca tcatcaccat caccaccacc accaccactg
     1321 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1381 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     1441 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     1501 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1561 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1621 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1681 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1741 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1801 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1861 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1921 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1981 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2041 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2101 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2161 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2221 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2281 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2341 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2401 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2461 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2521 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2581 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2641 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2701 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2761 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2821 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2881 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2941 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3001 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3061 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3241 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3841 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3901 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3961 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4021 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4081 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4141 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     4201 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4261 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     4321 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     4381 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     4441 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     4501 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     4561 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     4621 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     4681 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4741 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     4801 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     4861 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     4921 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     4981 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     5041 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     5101 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5161 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     5221 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     5281 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     5341 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     5401 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     5461 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     5521 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     5581 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     5641 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     5701 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     5761 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     5821 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     5881 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     5941 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     6001 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     6061 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     6121 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     6181 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     6241 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     6301 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     6361 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     6421 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     6481 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     6541 agaggatcga gatctcgatc ccgcgaaat
//
