LOCUS       sequence-354305-        6548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A011QK89.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6548)
  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 6548)
  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: A0A011QK89
FEATURES             Location/Qualifiers
     source          1..6548
                     /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             1268..1297
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1350..1368)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1364..1411
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1448..1903
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1535..1554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1745..1766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1929..2033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2034..2102,2103..2894)
                     /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(2252..2271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3065..3653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3554..3573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3807..3824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3839..3978
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3904..3923)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4064..4086
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4080..4271)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5080..6162)
                     /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     6124..6143
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6163..6240)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6446..6465
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggaaagcatt gaggcggttg ttattggtgc
      121 aggtgttgtg ggtctggcgt gtgcacgtga actggcgcgt cgtggttttg aaacggtaat
      181 tttagaacgc catggagcgt ttgggacgga gacgagtgct cgcaatagtg aagttattca
      241 tgcgggcttg tactatccca ctgactctct taaggctcgt ctctgtgttg cgggacgcca
      301 gcaactctat gcgttttgtg caacacatgc aatttctcac caacgctgtg ggaaattagt
      361 agtagcgact agtcctgccc aagaaagccg tttggccgcg ttgcaaaagc aaggtgaagc
      421 caacggtgtc gatgatctgc agcgtttatc ggcggcagaa gcacgtgcct tagagccagg
      481 tttggcatgc actgcagcat tgttatctcc gtcaacaggc attgttgact ctcatggtct
      541 tatgttggcc ctgcttggag atgcggagac tgcaggtgcg gcccttgccc tgcattcgcc
      601 tttactgcgt gggagcctgg atgcgaacac ccctggaatt gttctggaaa gtggtggggc
      661 cgacggctta cgctttaaag cccgtcgcgt tatcaacgct gcaggtctgt gggctccgca
      721 agtggcagca agtttagccg gattcccacg tacactgatc cctgcgaatt tccacgccaa
      781 gggatcctat tatgcgctta ccgggcgcac accgttttca cgtttggttt atccgcttcc
      841 agaagcaggt ggcttagggg tgcacctcac gcttgatctt ggcggtcagg cccgttttgg
      901 cccggacgtt gaatggctgc ctgatccgac acccggtcaa ccgattgacg aaccagatta
      961 tcgtgtggat ccggcacgtg cggatgcatt ctatgcagag attcgtcgtt attggccggc
     1021 tttgccggat gccgccctga ctccggccta tgcgggaatt cgtccaaaga ttgtgggtcc
     1081 aggagcgcca gcagcggatt tcttaattca gggaccagca caacatggca ttgccggatt
     1141 agtaaatttg tttggtattg aatcacctgg gttaacagcg tgcctggcga ttgccgaacg
     1201 tgccgcggat gccgcggacg gtacgcgcga acgtcaattc cgtgcacatg gcgcggccgc
     1261 actcgagcat catcaccatc accaccacca ccaccactga gatccggctg ctaacaaagc
     1321 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1381 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     1441 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     1501 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     1561 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     1621 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     1681 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1741 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     1801 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1861 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     1921 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1981 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2041 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2101 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2161 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2221 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2281 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2341 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2401 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2461 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2521 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     2581 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2641 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2701 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2761 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2821 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2881 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2941 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3001 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3061 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3121 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3181 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3241 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3301 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3361 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3421 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3481 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3541 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3601 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3661 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3721 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3781 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3841 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     3901 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3961 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4021 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4081 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     4141 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     4201 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     4261 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     4321 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     4381 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     4441 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     4501 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     4561 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     4621 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     4681 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg
     4741 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     4801 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4861 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     4921 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     4981 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     5041 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5101 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5161 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     5221 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     5281 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     5341 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     5401 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     5461 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     5521 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     5581 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     5641 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     5701 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     5761 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     5821 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     5881 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     5941 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     6001 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     6061 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     6121 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     6181 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     6241 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     6301 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     6361 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6421 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6481 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     6541 cgcgaaat
//
