LOCUS       sequence-354433-        6491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier B9S0Y9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6491)
  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 6491)
  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: B9S0Y9
FEATURES             Location/Qualifiers
     source          1..6491
                     /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             1211..1240
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1293..1311)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1307..1354
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1391..1846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1478..1497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1688..1709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1872..1976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1977..2045,2046..2837)
                     /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(2195..2214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3008..3596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3497..3516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3750..3767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3782..3921
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3847..3866)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4007..4029
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4023..4214)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5023..6105)
                     /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     6067..6086
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6106..6183)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6389..6408
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggagattacc aatgttatgg aatacgaaga
      121 aatcgcaaga cagaaattgc cgaaaatggt ttacgactac tatgcttccg gcgccgagga
      181 tcagtggacc cttaaagaaa acagaaatgc attctcaagg atcttgttca ggccacgtat
      241 tctaatcgat gtgtctaaga ttgatatgac aactagtgtc ctaggtttta agatatctat
      301 gcctatcatg atcgctccga cagccatgca gaagatggct catccggaag gcgaatacgc
      361 aacagccaga gctgccagcg ccgctggaac gataatgacc ctgagtagtt gggccacttc
      421 tagtgttgaa gaggtagcat caactggccc cggtatcaga ttctttcagc tgtatgttta
      481 tagagatagg aacgttgttg ctcagttggt gagaagagcc gagagagccg gcttcaaggc
      541 aatagcccta accgttgata cacccagact gggtcgtaga gaggccgata tcaagaatcg
      601 tttcaccctt ccacccttct tgactttgaa gaactttgaa ggcctagacc taggcaaaat
      661 ggataagtca gacgattcgg gcctctcgtc ctacgttgct ggccaaatag acagaactct
      721 tagttggaag gatattaaat ggttacaaac tatcaccagt ttgccgatat tagttaaagg
      781 ggtcttgact gcagaagata cgagacttgc tattcaaaac ggggccgcag gcataattgt
      841 ttctaatcat ggagctagac agctagacta cgtgcctgct accatcatgg ctttagagga
      901 agtagtgaag gcagctcagg gtcgagtccc tgtatttctt gacggtggag tgagaagagg
      961 tactgacgta ttcaaagcat tggcccttgg tgcttctgga attttcatag gccgtcctgt
     1021 ggtcttctca ctcgcagccg agggtgaggc cggtatccgt aaagttctgc agatgctgag
     1081 agatgagttc gagctgacca tggcattaag tggatgccga tctttgcgag agatcacacg
     1141 agaccacata gttacagact gggatctacc taggcctgta gcctctgcaa gattggctgc
     1201 agctcttgag caccaccatc accaccacca ccaccaccac tgagatccgg ctgctaacaa
     1261 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1321 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     1381 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1441 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1501 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1561 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1621 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1681 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1741 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     1801 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     1861 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1921 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1981 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2041 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2101 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2161 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2221 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2281 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2341 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2401 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2461 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2521 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2581 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2641 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2701 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2761 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2821 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2881 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2941 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3001 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3061 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3121 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3781 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     3841 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     3901 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3961 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4021 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     4081 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     4141 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     4201 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     4261 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     4321 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     4381 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     4441 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     4501 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     4561 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     4621 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     4681 ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     4741 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     4801 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     4861 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     4921 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     4981 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     5041 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5101 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     5161 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     5221 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     5281 atcatcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     5341 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     5401 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     5461 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     5521 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     5581 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     5641 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     5701 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     5761 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     5821 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     5881 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     5941 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     6001 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     6061 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     6121 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     6181 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     6241 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca
     6301 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     6361 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     6421 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga
     6481 tcccgcgaaa t
//
