LOCUS       sequence-354442-        6461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0A8UQ43.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6461)
  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 6461)
  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: A0A0A8UQ43
FEATURES             Location/Qualifiers
     source          1..6461
                     /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             1181..1210
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1263..1281)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1277..1324
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1361..1816
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1448..1467)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1658..1679
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1842..1946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1947..2015,2016..2807)
                     /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(2165..2184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2978..3566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3467..3486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3720..3737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3752..3891
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3817..3836)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3977..3999
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3993..4184)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4993..6075)
                     /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     6037..6056
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6076..6153)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6359..6378
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggacgagtac acgagcacac aatctttcta
      121 caagaaggct aagacagttc ttagtcccga cgttttctct ttcatcgacg gtggaagtct
      181 tgatgagctg gctctaaaga ggaacagaga gacattttct aaggttttga tttctcctcg
      241 tgttctgaga ggagtgaaca agtctaccac tcaagtgaaa ttgttcgacc acacacactc
      301 tgctccagtt ctaatagccc cagccgcata tcaatccttg ctgtcttcca agggcgagct
      361 agatatgctg gaagctgcca accaattcaa tacaactatg attctatcca tgttctcaag
      421 tattgattac actttgatcg cacgaaacaa gaaggtccca gtttggagcc agatgtattt
      481 tctcaaggac aggggaatca ataagaactt tatacaatgg atggaagaat taaactttga
      541 cgcacttgtt gttactgttg atgcaccagt ctatgcaaag aaagagagag aaaggtccca
      601 acctctcaga ttcccagaga acacctcttt cccacatctt aagaaaatcg ggatcccatt
      661 tgaagagtgt ttgaagacta aacgacatct ttccacgctg ttagaccatt caatttcatg
      721 ggacgacttg gactggctag ctggggtgac gaagctccct atcatcctaa aaggaataat
      781 tgatccaaga gacactgaga tagcactcac atatcccaac gttaaaggaa tcgtgatctc
      841 gaatcacggt ggccgacagt tagatagctc cattgctcct cttgaggtga tgcacaaaca
      901 cagagagaag gccgggaata aagtcaagct cttcttggat ggtggcatcg cacgtgggtc
      961 tgacatattc aaagctctcg ctttgggtgc agatgcaacc ttgactggga gaactgcttt
     1021 gtgggcatta tccatcggcg gctctgctgg ggttttccac gcattatcga tcttgcagca
     1081 ggagctccaa gaaactatga tcctttcagg ttgcagcacc ataaaggata ttacaagcga
     1141 ctacttggct tgtaaaggcg atacggcagc agctctggaa caccaccatc accaccacca
     1201 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1261 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1321 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1381 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1441 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1501 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1561 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1621 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1681 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1741 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1801 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1861 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1921 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1981 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2041 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2101 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2161 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2221 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2281 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2341 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2401 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2461 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2521 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2581 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2641 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2701 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2761 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2821 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2881 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2941 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3001 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3061 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3121 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3181 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3241 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3301 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3361 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3421 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3481 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3541 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3601 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3661 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3721 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3781 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     3841 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3901 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3961 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4021 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4081 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4141 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4201 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4261 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4321 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4381 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4441 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4501 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4561 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4621 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     4681 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     4741 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     4801 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     4861 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     4921 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     4981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     5101 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     5161 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     5221 cggcgggata taacatgagc tgtcttcggt atcatcgtat cccactaccg agatatccgc
     5281 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     5341 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     5401 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     5461 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     5521 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     5581 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     5641 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     5701 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     5761 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     5821 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     5881 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     5941 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     6001 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     6061 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     6121 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     6181 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6241 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6301 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6361 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6421 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa t
//
