LOCUS       sequence-354405-        6944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0P7Z009.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  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 6944)
  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: A0A0P7Z009
FEATURES             Location/Qualifiers
     source          1..6944
                     /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             1664..1693
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1746..1764)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1760..1807
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1844..2299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1931..1950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2141..2162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2325..2429
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2430..2498,2499..3290)
                     /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(2648..2667)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3461..4049
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3950..3969
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4203..4220
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4235..4374
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4300..4319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4460..4482
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4476..4667)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5476..6558)
                     /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     6520..6539
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6559..6636)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6842..6861
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggcaacctta gcatccgatc acccgttggc
      121 accggtacgt caagcagtag gtgatgcccc tttactgaca gacgccgatt cattagccct
      181 ctattctcat gatgtgctgg caaaaggcgc cgagccttta gcagtatttc gtccggattc
      241 cgtggaagca ttagctgctg gcattggcgc ggctacaggt gcagggatgg caattgtccc
      301 acgtggcggt ggaatgagct atactggtgg ttacttatat tcaggcggtc cgttcttgtt
      361 agtagatacc gctgcgctgg atcgcgttat tgaggttaat gagacagata tgaccgtgac
      421 cgttgaagca gggatttcat gggacgcatt acgtcgtgcc ctggcaccga aaggtttacg
      481 tgtaatggca tggggaaccc tttctggcat ccgcgcgagt gtgggcggtg gaatgagcca
      541 aaacggggtt ttctggggag cggcccgtgg gacagctgca gactcagcaa tttcgtttga
      601 tgtagtgctg gccgacggca cagtgatgtc gaccggctcg gacttcttcc gtccgtacgg
      661 cccagacttg accggcttgt ttgcggcaga ctgtggtgcc tttggagtta aagcccgcgt
      721 gaccttacgc ctcgttcgcg aggcctcagc ctttgcgtac ggtagctttt catttggcag
      781 tcacgcagct ctcttaggtg cgatgtcgga aattgcccgc gcggaactgg catctgaaag
      841 ttttgctttt gatccttttc tccaagccca gcgtatgaaa cgcgacactc tcgtgaaaga
      901 cgccaaacaa ttaggtaata tgttgaagag ccaggcgaaa agcggtggag tcttcaaagc
      961 cctgaaggaa ggcgcaaaag tggctatggc aggacgttcg ttcctggacg gtgtcccgtt
     1021 cagcttacat tgcattgcgg aaggccgtca ccaaggggcg gtggacgcag atatgaagcg
     1081 catcgcggag attgttcgcg aacaaggtgg tgagtcgatt gaaaactcga ttccgaagat
     1141 tttacgtgca aatcctttcc cacccccgaa ttctatgtta ggtccggaag gcgagcgttg
     1201 ggtacccgtc catggctttc tgccgcactc gaaactcgtc ccagcatggg agcgcttaga
     1261 ggcactgtgg gccgcaaatg cgggcgaaat ggaacgtctg aaagtcgaaa ccggcgccct
     1321 cattgcagca acaggccgta ctagctgctt gatcgagccg gtgttcttct ggcctggcga
     1381 acacaatccg ttgcaccgcc acgcggttga ggcagatcac ctggcaaagt taacatctcg
     1441 tcccgacagt cccgatgcaa atgctttagt tgatcgtctg aagggtgagg tcatcgctat
     1501 ctttcgtgag cttggagcta ttcacttaca agtggcgcgt acgtatccgc ttcgccaatc
     1561 tcatgatccg caaggatggg caatgctcga agcgctgaaa cgccaagtgg acccgaaagg
     1621 tctcatgaat cccggctccc tcggcctggc ggccgcactc gagcatcatc accatcacca
     1681 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1741 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1801 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     1861 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1921 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1981 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2041 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2101 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2161 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2221 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2281 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2341 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2401 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2461 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2521 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2581 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2641 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2701 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2761 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2821 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2881 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2941 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     3001 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3061 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3121 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3181 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3241 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3301 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3361 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3421 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3481 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3541 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3601 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3661 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3721 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3781 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3841 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3901 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3961 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4021 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4081 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4141 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4201 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     4261 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     4321 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4381 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4441 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4501 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     4561 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     4621 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     4681 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     4741 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     4801 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     4861 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     4921 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4981 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     5041 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     5101 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     5161 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     5221 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     5281 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     5341 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     5401 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     5461 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5521 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5581 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5641 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5701 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     5761 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5821 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     5881 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5941 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     6001 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     6061 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     6121 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     6181 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     6241 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     6301 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     6361 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     6421 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     6481 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     6541 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     6601 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     6661 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     6721 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6781 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6841 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6901 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaat
//
