LOCUS       sequence-354315-        6500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier D8N7C7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6500)
  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 6500)
  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: D8N7C7
FEATURES             Location/Qualifiers
     source          1..6500
                     /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             1220..1249
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1302..1320)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1316..1363
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1400..1855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1487..1506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1697..1718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1881..1985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1986..2054,2055..2846)
                     /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(2204..2223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3017..3605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3506..3525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3759..3776
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3791..3930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3856..3875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4016..4038
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4032..4223)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5032..6114)
                     /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     6076..6095
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6115..6192)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6398..6417
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gactgcactc gacccggcct tagcacgtct
      121 gcgtgatgct atcctgcaag ccgcagctga ccgtgcacct ctgactttgc gtggtggtgg
      181 cacgaaagac ttttatggcc gtcgtccggc cccgggtgcg accgttttgg atacgcgtga
      241 atggtcgggt attgtggact atgactgcgc ggagcttgtc gtaactgctc gttgtggaac
      301 ccctttagca gaggtagaag cagcgttagc agaacatcgc caaatgttag catttgaacc
      361 gccacatttt gccgctggtg gcgcccgtgc aactgtaggt ggagcttttg cctgcggcct
      421 gtcaggtccg cgtcgtcaaa gcgttggcgc attgcgcgac tttgttttag gtgcggtaat
      481 tttggatgga cgtggtgatc tgctgaactt tggtggtcag gtcatgaaga acgtagccgg
      541 ctatgatgtt agccgtttaa tggcgggtgc tttgggaacc ttgggcttag tcgctcaagt
      601 gtcattaaaa gttcttccta tgcccttctg cgatgctact ttacgttttg caatgcctca
      661 ggctgaggct attgacactc tgaatcgttg gggtgggcaa ccactcccga tcgcggcaag
      721 tgcgtggatt gacggaacct tatgggtacg cctttgtggt gctgatgctg ctgttcgtgc
      781 ggcgcgcgca cgtttgggcg gcgatcgcgt cgacgaagca gaagccttag ctttgtggcg
      841 tgatttgcgt gaacaaagcc ataccttctt cgcacgtgca gtgcaaggaa gtcttccatt
      901 atggcgcatt gcgttgccgc ctgcaacgcc gcccttagct ttgtcgagtg cagcggcgga
      961 tgatcaactc atcgagtggg gcggtggtca acgttggtgg atcgatactg gcaacactcc
     1021 ggcagatgca attcgtgatg cagtacgcca agcaggtgga catgccacac tgttccgcaa
     1081 tggagatcgt gacagtgcgg ttttcacgcc cgtggcagct cctctgatgg cagttcataa
     1141 acgcctgaag gattcctttg acccgcatgg tatctttaat cctggacgtc tttatgcgga
     1201 ttttgcggcc gcactcgagc atcatcacca tcaccaccac caccaccact gagatccggc
     1261 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1321 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     1381 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1441 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1501 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1561 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1621 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1681 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     1741 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     1801 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     1861 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1921 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1981 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2041 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2101 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2161 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2221 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2281 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2341 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2401 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2461 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2521 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2581 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2641 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2701 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2761 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2821 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2881 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2941 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3001 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3061 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3781 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3841 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     3901 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3961 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4021 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     4081 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4141 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4201 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4261 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4321 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4381 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     4441 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     4501 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     4561 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     4621 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     4681 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
     4741 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
     4801 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     4861 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     4921 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     4981 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     5041 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5101 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     5161 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     5221 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     5281 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     5341 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     5401 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     5461 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     5521 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     5581 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     5641 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     5701 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     5761 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     5821 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     5881 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     5941 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     6001 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     6061 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     6121 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     6181 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     6241 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     6301 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     6361 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     6421 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     6481 agatctcgat cccgcgaaat
//
