LOCUS       sequence-348048-        6641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier F0TBY7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6641)
  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 6641)
  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: F0TBY7
FEATURES             Location/Qualifiers
     source          1..6641
                     /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             1361..1390
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1443..1461)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1457..1504
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1541..1996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1628..1647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1838..1859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2022..2126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2127..2195,2196..2987)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAALSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3158..3746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3647..3666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3900..3917
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3932..4071
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3997..4016)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4157..4179
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4173..4364)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5173..6255)
                     /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     6217..6236
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6256..6333)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6539..6558
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gagatacatc tgttcatact gtaatatata
      121 tatctacgac gaagaaaagg gcgacttgga aactaagctc gaacccggta cgaccctaga
      181 ggaaattccc gacagctggt tgtgcccggt gtgcggcatg ccaaaggagt atttaaagcc
      241 agtcaataat aagatatttg agataaaagc caagtcttac accgagactc aacaccagac
      301 tcaagacatc aactattatc gatcgatcgc taggaatatg ttacacggaa tctgcggaga
      361 atatgacgta tgtaatggga agccagatag aatgtgctct ggacagaact ttggcggtag
      421 catcggattt ggcggcgctg gacaagggaa gacatttgaa gaaaattaca actctctaca
      481 gagatataaa cttaagatga gggtcatcaa ggaccataaa gagcccgaaa tgtccttgag
      541 catatttaac aaacagattg ctttgcctgt tatgggatct tccctctctg gcgtcaagaa
      601 ctctatgaat gattcaatcc cagaggagac tttctaccga ggactattgc atggcgcaat
      661 gagctcaggt acaattggta tggtaggtaa cactaacgac gtgcctgatg atctcggtgt
      721 taagactgtc ggtgagaatc acggctgggg aatacctatc tttaagccac aatctcagga
      781 aaggctattg gaactcatca agcaagctga ggaactgaac gtcttagctg ctggagtaga
      841 ccttgagggt gcagggagta ctttctggaa gactgcaaag aaaccagtct acagaaaggg
      901 tgaaaacgac ttaatcgagc tcgttgatag tacggaactt cctttgatct tcaagggaat
      961 catgtgccgt gaagatgctg ctaagctact cgatgttggc gctgctgcct gttacgtcag
     1021 taatcacggt ggtcgagtct tggatggagc tcagggagtt gccgaggttc taccagaaat
     1081 atcaagtgaa gtcgacggaa agattcctgt gcttgcagac ggtgctattc gaacaggtta
     1141 cgacgttctg aaaattttgg ctctgggagc tgacgtcgct ctgattggca gacctctggc
     1201 cagattgtct ctggctggtg gagaggttcc ggtgaaactt tactacaaat acgtcaagga
     1261 tgatttgcgt aatgctatgt tgttaacagg ttgtgataat ctcaacgaaa tcaacatgga
     1321 tattctcact ggcccgtcga tgaacgctgc tgctttggaa caccaccatc accaccacca
     1381 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1441 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1501 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1561 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1621 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1681 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1741 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1801 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1861 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1921 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1981 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2041 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2101 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2161 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2221 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2281 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2341 ttctgctatg tggcgcggca ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2401 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2461 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2521 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2581 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2641 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2701 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2761 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2821 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2881 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2941 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3001 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3061 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3121 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3181 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3241 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3301 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3361 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3421 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3481 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3541 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3601 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3661 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3721 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3781 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3841 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3901 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3961 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4021 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4081 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4141 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4201 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4261 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4321 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4381 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4441 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4501 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4561 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4621 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4681 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4741 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4801 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     4861 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     4921 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     4981 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     5041 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5101 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     5161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     5281 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     5341 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     5401 cggcgggata taacatgagc tgtcttcggt atcatcgtat cccactaccg agatatccgc
     5461 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     5521 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     5581 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     5641 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     5701 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     5761 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     5821 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     5881 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     5941 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     6001 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     6061 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     6121 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     6181 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     6241 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     6301 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     6361 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6421 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6481 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6541 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6601 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa t
//
