LOCUS       sequence-354460-        6614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0K2AU21.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6614)
  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 6614)
  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: A0A0K2AU21
FEATURES             Location/Qualifiers
     source          1..6614
                     /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             1334..1363
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1416..1434)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1430..1477
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1514..1969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1601..1620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1811..1832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1995..2099
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2100..2168,2169..2960)
                     /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(2318..2337)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3131..3719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3620..3639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3873..3890
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3905..4044
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3970..3989)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4130..4152
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4146..4337)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5146..6228)
                     /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     6190..6209
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6229..6306)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6512..6531
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggaacgtgtt cgtaccgctg tagcccgtca
      121 tgtcgcctat gattgtgacg ttctggttat tggcggcggt atcgttggct taagtaccgc
      181 ctatgcaatt acacgtgcag cgcctggaac tcgcgtaaca gttttggaga aagagccggg
      241 accagcacgt caccaaacag gtcgtaattc gggagtgatc catagcggag tatattaccg
      301 cccaggttcg ttaaaagcac gttatgccgt ccgtggttgt gcggaaatgg ttaaattctg
      361 tgctgaatat ggtattgctc atgccgttac tggtaaactt attgtggcaa ctcaccgtag
      421 tgaacttcct cgtcttcacg cgcttgtaca acgtggtcgc gaaaacggta tccctgttcg
      481 tgaacttgga gcagcccaaa ttcctgaata tgaacctgaa gtacgtggtt tagcagctat
      541 tcatgtaggc accactggtg tgtgtgatta tgtagccgta gcacgccagc ttgcgcgcgc
      601 ctctggcgca gaaatccgtt atggggccca agttcgccat attgaccgtc gtcctggcct
      661 tggcgtggca gtgcgtacgc gttccggaga tgtacttcgt gcccgtgttt tagtgaattg
      721 tgcgggattg cattgtgatg agatcgcccg tgccacagga gatgacccag gcatgcgtat
      781 cgttccattt cgcggtgaat attatgaact tgcgcgtccg gaattggtac gcggtctcgt
      841 atatcccgtc ccagatccag cgttcccctt tctgggtgtt catttaacac gtggaatcga
      901 tgggtctgtg cacgtcggac ccaatgcagt ccctgcactg gcgcgcgaag gttacggctg
      961 ggatgtagtt cgtccgcgtg aattaggcgc aacactggcc tggccgggct cgtggcaaat
     1021 tgctcgtcgt cactggcgtt atggagccgg ggagcttcgc cgtagtctgt cgaaaggtgc
     1081 gtttacggca gcagtgcgtc gtcttcttcc ggccgtgaca gatggtgact tagtcccgtc
     1141 tccagcgggc gtacgcgccc aagcggtttt agcagatgga actctggtgg atgatttcct
     1201 ggtccgcgaa ggcgcacacg cggttcatgt tcttaacgca ccaagcccag cggcgactgc
     1261 cgcattaccg attggccgcg aagtcggtcg ccgtgcatta gctgcactgg gagattcagc
     1321 ggccgcactc gagcatcatc accatcacca ccaccaccac cactgagatc cggctgctaa
     1381 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1441 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1501 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1561 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1621 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1681 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1741 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1801 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     1861 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1921 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     1981 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2041 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2101 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2161 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2221 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2281 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2341 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2401 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2461 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2521 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2581 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2641 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2701 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2761 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2821 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2881 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2941 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3001 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3061 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3241 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3301 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3361 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3421 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3481 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3541 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3601 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3661 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3721 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3781 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3841 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3901 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     3961 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     4021 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     4081 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4141 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     4201 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     4261 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     4321 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     4381 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     4441 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     4501 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     4561 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     4621 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     4681 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     4741 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     4801 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     4861 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     4921 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     4981 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     5041 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     5101 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     5161 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5221 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     5281 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5341 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     5401 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     5461 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     5521 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     5581 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     5641 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     5701 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     5761 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     5821 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     5881 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     5941 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     6001 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     6061 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     6121 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     6181 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     6241 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     6301 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     6361 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     6421 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     6481 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     6541 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     6601 cgatcccgcg aaat
//
