LOCUS       sequence-354446-        6557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier A0A0N1ND03.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6557)
  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 6557)
  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: A0A0N1ND03
FEATURES             Location/Qualifiers
     source          1..6557
                     /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             1277..1306
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1359..1377)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1373..1420
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1457..1912
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1544..1563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1754..1775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1938..2042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2043..2111,2112..2903)
                     /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(2261..2280)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3074..3662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3563..3582
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3816..3833
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3848..3987
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3913..3932)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4073..4095
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4089..4280)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5089..6171)
                     /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     6133..6152
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6172..6249)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6455..6474
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcctaggaat agagctgcta gaatcatttc
      121 cgttgctgat gcccagagga gagctagaag ggttataccg agacttttga cggattacat
      181 agaaggcggg gctgaggatg aactaaccct agctgaaaat gttcgtgcat tccaggaatt
      241 gggtatccgt cccagaatgg gtgtagactt cgaaccagat attaccacca ctgtcctggg
      301 tacgagggtt gaattacccg ttcttttagc tccaagtggg atgatccagt taatacatcc
      361 tgacggtggt gttggtgtgg ctagggctgc tgccgcagca ggcaccattt ccgttctctc
      421 tagaaccgca atgtgctccc cacgtgaggt tgccgctgga gcacccggtc cgcactggtt
      481 ccagatcact agtgctggcg gtcgtgatgc tgtgcgtaga ttgatggaca ctgccgctga
      541 agctggtttc caaggacttg tcgtaaccat ggacggacca cccccgggaa accgagaagg
      601 agagctgaga cacggagtcg ccccgcctgt gaggccgagc aagagacttg ccgctcacat
      661 tgcaggtcag atgttggcca ggcccaggtg gttggcctct atggcccctt cactgagaag
      721 acagttggct gccttgagtg gagcaagcag tcttgtgagt tcgggtgtac tgcacggtag
      781 cgcacgtttt acttgggctg atgttgaatg gatgaaaagg gaatgggccg gacatgttct
      841 tgttaagggt gtcttgactg gaagtgatgc cttggccgct agagatagtg gggcagatgc
      901 aattatcgtg tctaaccacg gtggtagatc tctggatggc gttcctgcta caattcgtgc
      961 tctaccagag gtcgttgccg ctgtaggacc aactactgag gtattcttgg atggtggggt
     1021 tagacgagca acctgtgttg tgaaagcact atcattagga gcaagagccg tatttgtggg
     1081 cagaccattt gtatatggct tggctgctgc cggggaacca ggtgtggccc gaatcttgga
     1141 tctgttccga tccgagttgg taaggactct tagactaatg gggtgccccg gcgtcgccga
     1201 ccttaatcca gggtggcttc aactaccacc acctagagag gtcccagcag cactaccaca
     1261 ggcagcagcc ttggaacacc accatcacca ccaccaccac caccactgag atccggctgc
     1321 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     1381 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1441 cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1501 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1561 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1621 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1681 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1741 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1801 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1861 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     1921 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1981 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2041 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2101 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2161 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2221 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2281 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2341 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2401 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2461 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2521 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2581 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2641 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2701 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2761 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2821 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2881 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2941 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3001 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3061 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3121 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3181 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3241 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3301 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3361 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3421 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3481 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3541 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3601 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3661 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3721 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3781 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3841 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     3901 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3961 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     4021 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4081 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     4141 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     4201 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     4261 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     4321 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     4381 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     4441 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     4501 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     4561 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     4621 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     4681 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     4741 acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     4801 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     4861 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     4921 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     4981 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     5041 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     5101 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5161 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     5221 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     5281 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     5341 ttcggtatca tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     5401 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     5461 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     5521 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5581 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5641 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5701 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5761 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5821 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5881 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5941 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     6001 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     6061 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     6121 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     6181 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     6241 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     6301 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     6361 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     6421 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     6481 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     6541 tctcgatccc gcgaaat
//
