LOCUS       sequence-354353-        6752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier U2ENB5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6752)
  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 6752)
  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: U2ENB5
FEATURES             Location/Qualifiers
     source          1..6752
                     /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             1472..1501
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1554..1572)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1568..1615
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1652..2107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1739..1758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1949..1970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2133..2237
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2238..2306,2307..3098)
                     /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(2456..2475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3269..3857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3758..3777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4011..4028
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4043..4182
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4108..4127)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4268..4290
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4284..4475)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5284..6366)
                     /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     6328..6347
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6367..6444)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6650..6669
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gacccaagcc gtggacttca gcattgagac
      121 attttctagc gagctgggtg atgtcgaaat tattagcaac gaaaagcaac gcaaacgcct
      181 gtcaaaggat tttagctggt tctctccggt gttgaaggac ttcttcgaag gttatgaagc
      241 ggacgtagtc gtcaaaccta cgaccgaaga acaggtagcg caagtggtta agacctgtgt
      301 ggcgcactca gttcccctta cgctccgcgg tagtggcacc ggtaactacg gacagtcgac
      361 accgatgttt ggcggcgtgg ttatggacat gtcggcctac aaacagccgt tatggctgaa
      421 aggcggagtt gcgcgtatgc aggctgggat gcgcctggcg gaagtcaaca aatggaccgt
      481 gccagagggc tgggaaatgc gtaccatgtc gagcaccttt cgcttggcga cagtaggcgg
      541 gttcttcgcc ggcggatttg gtggcattgg ctcaatcact tacggccgtc tgggcgaacg
      601 cggcaacgtc ctgggcgcaa aggtgatgac cgttgaggcc gaaccgcaag taatcgaact
      661 gcgcggagat gatgccctgg acttgcatca tgcctatggc accaacggta ttctcttgga
      721 actggaattc gccatggcac cggcgttagc gtgggccgag gcaattgttg ttttcgacga
      781 ctttatggat gcggcgcgtt ttggccaggc gctgggagag gctggcggcg tcgtgaagaa
      841 gctggacacg gtcctcgccg acccgattcc tcaaatgatc gaattaggag attacgcgaa
      901 accgggacag cacgtcgcct tggtcattat tgccgaaaac gcgctgcagc ctatggcgga
      961 aatcgccgcc gagcataatg gtaccgtggc taaacaagta gccccggaag aagtttcaga
     1021 ttccaatact ttgatcgagt atacttggaa tcacaccacc ctgcgcgcct taaagcaata
     1081 caaagatgtg acgtacctgc agtcgggctt ccgtgcaggc aagaaccttg aacagattga
     1141 gcatatgtat caacactttg gcgatgaggt gccgatgcat ttggagttct tgaatattgg
     1201 cggcatgatg acctgtagtg gaatccaaat cgttcgttac accacccgcg aacgcttgga
     1261 cgaaattatt gcgtatcatg aggcgcacga cgtccatatc gccaatccgc acacgcacat
     1321 tctggaagat ggccataact ctggggaagc cgatccacgt cagcttgccc gcaagaaggc
     1381 ttatgatcca aaaggattgt tgaacccggg aaagatgcgc gcctggggtg aagtgaccga
     1441 gcttgacaac gttggcgcgg ccgcactcga gcatcatcac catcaccacc accaccacca
     1501 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1561 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1621 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     1681 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1741 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1801 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1861 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1921 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1981 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2041 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2101 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2161 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2221 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2281 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2341 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2401 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2461 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2521 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2581 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2641 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2701 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2761 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2821 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2881 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2941 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3001 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3061 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3121 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3181 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3241 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3301 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3361 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3421 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3481 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3541 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3601 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3661 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3721 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3781 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3841 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3901 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3961 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4021 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4081 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4141 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4201 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4261 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4321 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4381 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4441 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4501 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4561 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4621 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4681 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4741 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     4801 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     4861 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     4921 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     4981 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5041 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5101 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5161 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5221 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5281 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5341 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5401 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5461 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5521 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     5581 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5641 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5701 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5761 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     5821 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     5881 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     5941 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6001 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6061 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6121 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6181 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6241 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6301 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6361 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6421 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6481 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6541 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6601 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6661 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6721 cgtagaggat cgagatctcg atcccgcgaa at
//
