LOCUS       sequence-354321-        6821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier B9NTK7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6821)
  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 6821)
  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: B9NTK7
FEATURES             Location/Qualifiers
     source          1..6821
                     /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             1541..1570
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1623..1641)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1637..1684
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1721..2176
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1808..1827)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2018..2039
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2202..2306
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2307..2375,2376..3167)
                     /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(2525..2544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3338..3926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3827..3846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4080..4097
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4112..4251
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4177..4196)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4337..4359
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4353..4544)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5353..6435)
                     /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     6397..6416
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6436..6513)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6719..6738
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggaaatgcct aaaccatcac ccgacgtgct
      121 gcgccgtaaa ggggcggttg tagcacgctt gagccaagtt ctgccgcagg atgccattat
      181 ccaggatgaa gcggagacgc gcgcatacga gtgcgacgcg ctgactgcct ataagtgtgc
      241 gcccatgatc gcggttttac cctctactac tcaacaagtt tctgacgtat tacgtatttg
      301 tcacgaagag ggcgtgcctg ttgtcccacg cggagcagga acgagcctgg cgggtggagc
      361 gttaccgact gcagactgcg ttatcctggg cgttgctcgt atgaatgacg tgcttgaggt
      421 ggattatgat aaccgtttta ttcgtgtcca gacaggccgt accaatttgt cggtgaccgg
      481 ggcggtggaa gaagaggact tcttctatgc cccagatcct tcttcacagt tagcatgcgc
      541 tattgcgggg aatatcgcca tgaactcggg tggtgcccat tgcctgaaat acggggttac
      601 tactaacaac ttgatgggcg tgacaatggt catgatggac ggcacagttg ttgaaatcgg
      661 cggcgcgtat ctcgatgcag gtggcttgga tttgcttggt gtcatttgcg ggtcggaagg
      721 gcagcttggt gttgtaaccg aggccactct ccgtattctg cgcaagcctg aaggtgcgcg
      781 cccagtcctg atgggtttcg acgataacga ggttgccgga caatgtgtgt cggatatcat
      841 caaatcgggt attctcccgg tcgctatcga gtttatggat cgcctttgca ttcaggcctc
      901 ggaaaagttc gcacaagccg ggtaccctga ttgcgaggct cttctgatcg tagaggtcga
      961 gggctctgag gcggaaatcg actcgcagtt atcaaagatt gtggaaattg cgaagcgcca
     1021 taacccggtg gagctgcgtg agtcgcgttc tgccgaggag tcagcgcgca tctggcttgg
     1081 acgtaagtcg gctttcggtg cgatgggaca gctgaatgac tacatgtgcc tcgatgggac
     1141 gattccagtg tcatcactgc cgtttgttct gcgtcgtatt ggcgagctgt ccaaagatta
     1201 cggtttaccg gtgggaaacg tatttcatgc aggagacggg aatatgcatc ccctcatctt
     1261 atttgatgcg aacaagccgg gcgatttaga gaaatgcgag gccttcggtg ccgatattct
     1321 tactttgtgc gtagatgccg gtggatgtct gactggggag cacggagtcg gcattgaaaa
     1381 gcgcgacctg atgacgcatc agtatacgcc ccaagattta gaggcccaga tggccgtaaa
     1441 agacgttttc gatccgggct ggcttttaaa tccggccaag gtgtttccgc tggcggcgtc
     1501 agagccacgt cgtatggccg cggaagccgc agcactcgag catcatcacc atcaccacca
     1561 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1621 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1681 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1741 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1801 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1861 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1921 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1981 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2041 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2101 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2161 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2221 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2281 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2341 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2401 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2461 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2521 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2581 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2641 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2701 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2761 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2821 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2881 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2941 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3001 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3061 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3121 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3181 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3241 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3301 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3421 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3481 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3541 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3601 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3661 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3721 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3781 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3841 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3901 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3961 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4021 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4081 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4141 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4201 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4261 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4321 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4381 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4441 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4501 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4561 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4621 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4681 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4741 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4801 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4861 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4921 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4981 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     5041 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     5101 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     5161 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     5221 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5281 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     5341 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5401 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     5461 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     5521 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     5581 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     5641 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     5701 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     5761 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     5821 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     5881 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     5941 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     6001 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     6061 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     6121 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     6181 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     6241 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     6301 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     6361 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     6421 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     6481 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     6541 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     6601 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     6661 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     6721 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     6781 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa t
//
