LOCUS       sequence-354431-        6479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of UniProt identifier F4G5A4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6479)
  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 6479)
  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: F4G5A4
FEATURES             Location/Qualifiers
     source          1..6479
                     /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             1199..1228
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1281..1299)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1295..1342
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1379..1834
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1466..1485)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1676..1697
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1860..1964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1965..2033,2034..2825)
                     /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(2183..2202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2996..3584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3485..3504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3738..3755
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3770..3909
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3835..3854)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3995..4017
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4011..4202)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5011..6093)
                     /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     6055..6074
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6094..6171)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6377..6396
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtctccttca gatcgtatac ccccgggagt
      121 gtggaatgca attgattatg agagattagc tccacaagca atggatgccg gacgtcacgc
      181 atacgttgct ggcggttgtg ggtgggacgc cacagttgca gctaaccgtg ctgcatttgc
      241 aggctgggcc gttcttccac gacttttgcg agatgttcgt gccggacaca ccagattgca
      301 gttggcaggt atggatttgc ctcatcctct tcttttggcc cccgtagctc atcagcgtct
      361 ggcccacccg gatgctgaaa tagctaccgc tagagccgcc caagccactg gttcttgctt
      421 ggtagcttca actctgtctt cctgtactct tgaagacata gctgcagctt ctggtccagc
      481 aagatggttc cagctgtatt tgcagccgga acgagagcac agtctggatc tcctacgtag
      541 agccgaagct gccggttaca gagccattgt cctaacgtta gatgcctcaa ttcagttggc
      601 ctctaggggc gctttgcagg ccggttttgc tatgcctgct gactgcgtca gtgctaactt
      661 agccagatat cctcaacccg cccctgcaca gccagcagct ggtgagtcca gaattttcca
      721 aggtgccatg agacatgcac cacgttggga tgatcttcgt tggctactcg caagtactcg
      781 actgcccgtc tggattaaag gggtgcttca cccagaagat gccagagagt tacaagcagc
      841 tggggctgcc ggtctaatcg tctctaacca cggcggtcgt agcttggacg gagcaccagc
      901 ttctctccga atgctgccag ctctaagaac cgctgtgggt gctggctatc cactactctt
      961 agacggtggt gtcagatccg gccaggatgc ttttaaggct ctagcattgg gtgctgatgc
     1021 tgtgttggtt ggacgtcttc aggtgtacgc actcgctgtc gccggtgctt taggcgtcgc
     1081 tcacatgttg cagatgttgg tggaagagct tcacgcttgc atggcccagg ctggatgtgc
     1141 taggttgtca gatattacac atgatacctt aactccctca tgtgcagcag ccttagaaca
     1201 ccaccatcac caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     1261 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1321 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     1381 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1441 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1501 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1561 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1621 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1681 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1741 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1801 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1861 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1921 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1981 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2041 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2101 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2161 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2221 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2281 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2341 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2401 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2461 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     2521 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2581 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2641 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2701 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2761 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2821 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3121 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3421 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3481 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3541 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3601 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3661 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3721 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     3781 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     3841 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     3901 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3961 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4021 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     4081 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     4141 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     4201 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     4261 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     4321 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     4381 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     4441 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     4501 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4561 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     4621 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     4681 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     4741 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     4801 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     4861 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     4921 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     4981 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5041 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5101 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5161 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5221 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat catcgtatcc
     5281 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5341 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5401 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5461 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     5521 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     5581 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     5641 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     5701 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     5761 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     5821 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     5881 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     5941 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     6001 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     6061 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6121 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6181 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     6241 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6301 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6361 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     6421 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaat
//
