LOCUS       sequence-354304-        6800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A077SBA9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6800)
  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 6800)
  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: A0A077SBA9
FEATURES             Location/Qualifiers
     source          1..6800
                     /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             1520..1549
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1602..1620)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1616..1663
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1700..2155
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1787..1806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1997..2018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2181..2285
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2286..2354,2355..3146)
                     /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(2504..2523)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3317..3905
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3806..3825
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4059..4076
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4091..4230
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4156..4175)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4316..4338
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4332..4523)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5332..6414)
                     /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     6376..6395
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6415..6492)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6698..6717
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gaccgatcct cgtattctta gccttcagca
      121 agccgtccca gcactccgtc tgaaaactga gccagcagat cttgagcatt atggccgtga
      181 ctggacccgt cgctggacac ccaatccatt agcgattgcg cttccgggca gcgtcgaaga
      241 agtacaggcg gtcgttcgtt gggcaaatgc ccaagcggtt gcagtggtgc cgagtggcgg
      301 acgtactggg ttatcaggcg gagcggttgc tgctaacgga gaattagtgc tgtccctcga
      361 acgcttaaac aaaccgttag attttaatgc ggtggatcgc acattgacag tccaggcagg
      421 tatgcctctt gaagcagtcc ataatgctgc gcgcgaacaa ggtctcgtct atcctgttga
      481 cttcgctgca cgtggttcat gctctattgg tggtaacatt gccacaaacg ctggtggcat
      541 tcgcgtgatc cgctacggca acacgcgtga atgggtggcg ggcctgaaag tggtgacggg
      601 tagcggcgag ctgttagaac tgaacaacgc cctggttaag aatagcagcg gctacgactt
      661 ccgtcatctg atgattgggt cagaaggcac cctgggtatt gttgtcgagg cgactctgcg
      721 tttaaccgat ccgccaccgc cctcaaatgt gatgctttta gccctgccct cgttcgacgt
      781 gctcatgcaa gttttcgctg catttcgcgc tcaactgcgc ctcgaggcat ttgagttctt
      841 tacggatcgc gccttggaac atgttctggc gcacggcgcg caagcacctt ttgcagaaat
      901 ccacccgtac tatgtcgtta ccgaattcgc ggccggtgat gaagcgcaag aagccgcggc
      961 gatggctgcc tttgaaacgt gtatggagca aggttgggta agtgatggtg tgatttcgca
     1021 atcggacgct caagccgcgc aattgtggcg tttacgtgag ggaattacag aggcactggc
     1081 gcgctacacg ccatataaga acgacgtaag tgtgcgcatt tccgctatgc cagccttctt
     1141 agcggagacc caggctcttt tgcatgatgc atacccggat ttcgacgttg tctggtttgg
     1201 gcacattggc gacggcaacc tgcatattaa tgtgctgaaa ccagatgcga ccagccaagc
     1261 ggacttcgtg gccgcttgcg atcaagtaac taaattatta gcacaagccc tgcagcgctt
     1321 cgatggctcg atctctgccg aacacggcat tgggctggtg aagaaaagct acttgtggtc
     1381 gacccgttca gcggaagaaa tcgcgctgat gcgcggcatc aaacacgttc tggacccgca
     1441 cctgctgctg aatccgggta agcttttcga aacccacgat gcgccaacaa atatcccggc
     1501 aggcgcggcc gcactcgagc atcatcacca tcaccaccac caccaccact gagatccggc
     1561 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1621 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     1681 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1741 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1801 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1861 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1921 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1981 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2041 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2101 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     2161 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2281 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2341 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2401 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2461 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2521 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2581 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2641 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2701 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2761 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2821 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2881 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2941 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3001 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3061 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3121 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3181 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3241 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3301 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3361 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3421 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3481 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3541 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3601 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3661 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3721 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3781 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3841 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3901 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3961 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4021 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4081 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4141 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     4201 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     4261 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4321 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     4381 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4441 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4501 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4561 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4621 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4681 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     4741 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     4801 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     4861 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     4921 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     4981 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     5041 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     5101 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     5161 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     5221 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     5281 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     5341 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5401 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     5461 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     5521 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     5581 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     5641 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     5701 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     5761 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     5821 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     5881 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     5941 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     6001 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     6061 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     6121 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     6181 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     6241 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     6301 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     6361 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     6421 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     6481 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     6541 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     6601 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     6661 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     6721 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     6781 agatctcgat cccgcgaaat
//
