LOCUS       sequence-348029-        6917 bp    DNA     circular SYN 10-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0U5NXR6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6917)
  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 6917)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 10, 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: A0A0U5NXR6
FEATURES             Location/Qualifiers
     source          1..6917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(346..1428)
                     /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     1390..1409
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1429..1506)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1712..1731
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1815..1834
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1815..1833
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1834..1858
                     /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             1873..1895
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3451..3480
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(3533..3551)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3547..3594
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3631..4086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3718..3737)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3928..3949
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4112..4216
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4217..4285,4286..5077)
                     /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(4435..4454)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5248..5836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5737..5756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5990..6007
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6022..6161
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6087..6106)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6247..6269
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6263..6454)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
       61 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
      121 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
      181 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
      241 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
      301 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
      361 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      421 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
      481 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
      541 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
      601 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
      661 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
      721 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
      781 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
      841 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
      901 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
      961 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     1021 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     1081 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     1141 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     1201 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     1261 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     1321 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     1381 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     1441 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     1501 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     1561 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     1621 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     1681 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     1741 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     1801 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     1861 tctagaaata attttgttta actttaagaa ggagatatac atatgtcctc tcacctagga
     1921 caatggaaga cgattggacc caaatttaat gcattgcctg tccataccgc cttactacac
     1981 acagggaaag tcctggcttt cggtggctct gggaatgagc ctggtaggct taataattgg
     2041 tacgaacctg agatttatga gcctgactac acgggtagca ctgacggtac gataagaaag
     2101 atctcaaacg atggaattga aggtgatttg ttctgtgtgg gacacgcata tctgccggac
     2161 ggaaagttgt tcctggcagg tggtacgtat ttatacgacg gattaggaat agtgacggct
     2221 ccctttcccc cattctgggg tcttgatcac acttacactt ttgatccaga caccgagaaa
     2281 tttacccgat tgccgaatat gcgttacggt agatggtacc caacctgtgt tactatgggc
     2341 gacggcaaga ttgctacatt tgccggattg agcaagtggc caccctgggt ctggcagcct
     2401 gttaatgaaa tttatgacga acagaataac tcgtggcaga agattgaggg tggtaacaat
     2461 ctgatgccac tataccctcg tatgcacttg ttaccaaatg gtgacttgtt ttatgccgga
     2521 tcatataaca cccacatgac ttttccattc agtagatggg gctttaagat agcaactttc
     2581 tccttcgtag agaagaagtg gaagcattta gagcacccta aagatatgca caggcaagag
     2641 ggtagcactg ttctgttgcc tttagttaaa tccaagaatt acctggctga agtcatctta
     2701 ataggtggcg gagatatgaa tgcccatgca actcgaactt gcgaaaagat ttcattcaag
     2761 tccggaacca agcagaaata tatcccaacc aagtctatgg acaatgatag atactatgtg
     2821 tatcccgtaa ttcttccaga tcgaaatatt ctggttgtag gtggaaagtc gggtcacaat
     2881 agtcacagca ttaaacacaa atgtgaagag attgctcctg aagagataaa gcataatgac
     2941 aaggctattc ttgatactga gctgtataac accgaaactg aggagtggag taagcttgag
     3001 cctatgaaag tcgacagatt gtaccatgca aacgctatac ttttgcctga tggaaaagtc
     3061 atgacaatgg gatcaaaccc tacacgtaag tgtgtggaga agcgtattga gattttctcg
     3121 cccccgtact tgttcgcagg tgaccgacca gtcatcgacg atgttaaatc ggtgctgaag
     3181 tacgaggaat cgtttgagat tcagatgcat gacggcacag aaattctgga cactgctttg
     3241 atcaggccct ctaacaccac tcattgtctg aatccggagc agagatacat cgagatcgac
     3301 accgtccaaa atggagacaa gatttcatct aagatcccta agaatgagaa cttggtacca
     3361 ccaggatatt acatgctatt tgtccgtaat cactctggtg ttccatgcat cgctcctttt
     3421 gtaagaatca cacacgctgc tgctcttgag caccaccatc accaccacca ccaccaccac
     3481 tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     3541 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     3601 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     3661 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3721 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3781 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3841 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3901 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3961 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4021 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4081 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4141 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4201 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4261 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4321 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4381 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4441 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4501 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4561 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4621 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4681 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4741 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     4801 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4861 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4921 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4981 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5041 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5101 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5161 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5221 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5281 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5341 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     5401 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5461 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5521 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5581 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5641 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5701 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5761 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5821 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5881 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5941 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6001 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     6061 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6121 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     6181 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6241 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     6301 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     6361 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     6421 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     6481 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     6541 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     6601 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     6661 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     6721 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     6781 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     6841 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     6901 gcacgatcat gcgcacc
//
