LOCUS       sequence-344608-        6605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0F8B4A0.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6605)
  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 6605)
  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: A0A0F8B4A0
FEATURES             Location/Qualifiers
     source          1..6605
                     /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             1325..1354
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1407..1425)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1421..1468
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1505..1960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1592..1611)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1802..1823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1986..2090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2091..2159,2160..2951)
                     /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(2309..2328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3122..3710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3611..3630
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3864..3881
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3896..4035
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3961..3980)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4121..4143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4137..4328)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5137..6219)
                     /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     6181..6200
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6220..6297)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6503..6522
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gttattgtct agagctgtat tgttgagacc
      121 acgacgagtt ggactaagag gtttctctac ctctaggctc gcagctaggc aggctgattt
      181 cactcatgcc gttatcggcg ctggtgtagt tggacttgct atcgcccaaa ggcttgctac
      241 aatacaccca aagtcgagtg tgctgttatt ggaacgtcat accagccctg gaacggagac
      301 ttcttctcgt aattctgagg tcttgcatgc cggaatctat tatggtagga cctctctcaa
      361 ggctcagctc tgcgtgagag gtagacagct ggcttatgat ttctgtgccc gttatggcgt
      421 cgggcatgct cgagttggaa aatggatcgt cgcacaaaca ccagaacaac acgcagctct
      481 ggctggtttg gcttctactg ctgacgtgtt aggggttcca accagatgga tcagtgccga
      541 cgtagccaga gctgccgaac cactggttag agctgcagcc ggagtgttag aatctccgga
      601 gactggtata gtggacagcc atggtctaat ggttgcttta ttggggcagt ttgaggacgc
      661 cggtggtgtg ctggctcgag gcgcctgtgt agaaggagtg caggcagaga acggtggtat
      721 gggcggatgg actcttaagg ttcgagacgc tgctactggg gacatcgcag aagttgtggc
      781 cgagtcgatt gtaaatgccg caggattagg cgcagtgatg gttaacaact taatcgtcgc
      841 tcctgaacgt cgtagacaaa tgttttatgc taaaggtaac tattttgcct atgcaggtcc
      901 acagccacgt atcaagaggc tcatctatcc tgcaccaact cctggaggag gtggacttgg
      961 tactcacttg acgttggatc tcgcaggtcg tgtgaggttt gggccagacg tggaatgggt
     1021 tgatagactt aatgatatgg cagtcaactc ggccagactt ccagaggcca ttacagcaat
     1081 caagcagtac ttgcctggca ttgatgacaa cttgttagtt ccagactacg ccggtgtgcg
     1141 tcctaaattg gcccgagcag gagctgttgg agaaggaaag ggatttcagg atttcattat
     1201 tcagaaagag caaggttata ccggttgggt aaatttgtta ggtattgagt ctcctgggct
     1261 taccagttca ttggccattg ccgaaatggt tgaggggctc ctatacaagg cagcagcttt
     1321 ggaacaccac catcaccacc accaccacca ccactgagat ccggctgcta acaaagcccg
     1381 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1441 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     1501 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1561 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1621 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1681 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1741 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1801 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1861 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1921 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     1981 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2041 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2101 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2161 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2221 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2281 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2341 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2401 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2461 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2521 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2581 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     2641 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2701 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2761 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2821 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2881 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2941 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3001 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3061 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3121 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3181 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3241 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3301 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3361 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3421 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3481 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3541 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3601 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3661 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3721 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3781 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3841 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3901 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3961 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     4021 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4081 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4141 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     4201 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     4261 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4321 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4381 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4441 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     4501 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     4561 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     4621 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4681 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4741 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     4801 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4861 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4921 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4981 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     5041 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     5101 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5161 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5221 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5281 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5341 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcatc
     5401 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5461 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5521 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5581 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5641 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5701 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5761 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5821 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5881 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5941 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     6001 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6061 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6121 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     6181 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6241 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6301 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6361 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     6421 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6481 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6541 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     6601 gaaat
//
