LOCUS       sequence-305916-        8189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His6-TglA(C51), TglH, TglI and TglB in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8189)
  AUTHORS   Ting CP, Funk MA, Halaby SL, Zhang Z, Gonen T, van der Donk WA
  TITLE     Use of a scaffold peptide in the biosynthesis of amino acid-derived 
            natural products.
  JOURNAL   Science. 2019 Jul 19;365(6450):280-284. doi: 
            10.1126/science.aau6232.
  PUBMED    31320540
REFERENCE   2  (bases 1 to 8189)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2019-07-19- 19"; volume: "365"; 
            issue: "6450"; pages: "280-284"
COMMENT     Sequence Label: pRSFDuet1-tglA(C51)HIB
FEATURES             Location/Qualifiers
     source          1..8189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             78..95
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        4569..4587
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4588..4612
                     /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)."
     CDS             4721..4765
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4803..4821)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4817..4864
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5094..5909)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5094..5909)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5817..5836
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5910..6001)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6017..6766)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(6027..6044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6267..6286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6962..8044)
                     /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     8006..8025
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8045..8122)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        8168..8186
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(8187..8189,1..22)
                     /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)."
ORIGIN
        1 attgtgagcg gataacaatt cccctgtaga aataattttg tttaacttta ataaggagat
       61 ataccatggg cagcagccat caccatcatc accacagcca ggatccgatg ggacaaccca
      121 acgtgcagtc cgttgaaaac cagcaagcgt ccggcgatgt aaaagatctc gaaaacactc
      181 cgcaggccac agaggaggcg ttgtttgaag agtttgacct ggatgacatt gaagtcatcg
      241 agagcaaggt ctttgcctgc tgagtgcgcc atgccctctt gcgtgccgga ttgtagggat
      301 acgcaggagg gcttctttca tcagtgcgag tgttagccat gcctgatttc gtcaagcccg
      361 cgcctatcgg agtgggtatc cagtacaacc ccgaaatact ggactggttt ccattcgagg
      421 atatccaggt ggatattctg gagattttgc tcgataacat catggcgccc atggatggcc
      481 cgcaaattat caagccttct gcccaggcaa tgattgagcg gctagggcaa aaatttacct
      541 tgctggctca ctccaattac ggctgcgatt ttggtttcag tgccctcgaa gaaacagcgg
      601 cggtgcagcg ccatgtacct ctggcaaaaa tgctcaacag cccttgggtg gccaaccatt
      661 gtttctatgg ggatcagtcc tggctggata tctggagcag tcccattcag ttctccgcag
      721 ccgaagtggc gcgttgtgcc gatcgcgcac agtcgttgca gacgctttat ggcatgccgc
      781 tggctcatga aaatgccgcc tactaccttg aatgcccagg cgccgaaatg cgtgaggccg
      841 agtttctggc aagactggtg cagcgctcag ggacttttct tcaccttgac ctgcacaaca
      901 tttacaccaa ccacctcaat ctgaaaggtt tcgatctcaa ggactacatg gacacgctgc
      961 cgctggacaa ggtgatttcg gtgcatctgg ccggaggaag ctggcacggc ggcctgtatc
     1021 acgactggca tgacgcctgc gtgcccgagc cggtctggga cttgtacgaa gacctgcttt
     1081 cgcgtgcaca gccctctgct gtgattcttg agtatcaggg gcaggcgcat catgcccaga
     1141 ctaggatcat ggatgcgtcc gacgagtcga tgattgtcag ggatgtccag cgtgctcagg
     1201 caatctggag tcgttacaac aggcatccgc aggagcgtca gtatggaagc tgaacaggta
     1261 tggaaactgt ggcggcgcgt tctgcgcgat gagcgccttc aggcgcagtt gttttcggct
     1321 accgatgcca cgcactggtt gtcaggtttc agcgagtccg aaagcaagat cctctcggtg
     1381 tatgcgcagc agtttgaccg cgtcaaatgg ttcgtcgaga actatcagtt ccggctggtg
     1441 aactcttttt tgaacgcgct ggagaccggc gcgcccttgt cgttgcgggc gttgctgcac
     1501 ataaatgtgg acctgaatgc gcaatcgaag gccttcctgc gtgacaggca gtggcgcgat
     1561 tacgggccgc aggtctatac ctattgtgaa gatgtactgg gtttccttgc cgaggcggat
     1621 gagctgcagg gttatcctga aatccttgat ctgatgcggc tggagcgcga atcggttcgc
     1681 ctgtatcggg ggttggtcga tcctgagtcc ttgccggcag acaaccgtta tcagcgcacc
     1741 tcaatggctc gtctttacga gacacgtttt gccctgtctg gctggctcag acagaaggat
     1801 cagttgggcc tgacacgtct gcccgagagc accgagcatg tgctgatcta ccttcccacg
     1861 cttcaggctc gtcacaagtt cactttgatc aacgcgcagg ctgcccggct ttacaactgc
     1921 ctggagcaac ctcagtccgc tgccgggctg ttcatgctga tcaattcgga ctcggcgtcc
     1981 gtgccgggta gcgcagacct ggcgctgctc gacaggctgg aacagcttaa cgcgataagg
     2041 aagcctttgt agcctatgga aagctcacac tatttctggc ttcgctcgac cgggtttgcg
     2101 gtgcatcacc tgacccgctt ggggaaaatg gccgaattgc cgttgctgaa ggattttgaa
     2161 accgattatc gctccctgaa tactctacgc gacagcctgt tagagaagtc cataagccac
     2221 tcgtctcagg cctgtcgcaa gctgatccgc aagctgaatg aaaacctgcc gctgcaagtg
     2281 tctgatcttc cagaagcgct gcgtgatgac gcagtggatg aattggcgca ctggaacgag
     2341 cggcttggcc ggctcaagca acgtgtcgaa gtggatcagg aatatgccgt gtttctcgaa
     2401 agcgcgcggc aggccctgat tgatttcgtg aacgatgaag acgttgagca ggccgtgttc
     2461 atttccaacc cgaccgcgct gacccggctt cgggagctca ggcaagagcg ccatgcgaga
     2521 actgacagcc gcaagaagca gaaactgcgc ctggcctgga gctacgcaca gcgattctgt
     2581 tcaaaaaatg atacgtcgag ttttttcggc ccgctggcct gggggcggtt tgatcgtacg
     2641 caggtcgagc atgttcgtat cacgcagcac ggacccggct ggatcagaga acgccatacc
     2701 tttttcgaga gctgggttgt acagcggctg gtcgagcagt tgaatcgcca ttgcgttgat
     2761 gtgcagttca tgccgttgca actcaacccg ggctgttttt tgaatcaaga cacgttgcat
     2821 ctgcccgtca acaaacgtcg tcaagtcagt gccttgacgg ccagagtgct gggttacatt
     2881 cagtgccaga gcgctgtggc accgaccctg ttcgggttgc aggccgagtt gcaggatgtc
     2941 agcgccgggc aattgcgcga cctgatcgat catctggtcg cccagcagat tgtacgtcgc
     3001 ggctggcaga tttcccccag ggaacgaaag cctgttcagg tgctgcacac ttttctggcc
     3061 gacgcgcgcc tgtctgatga ctttcgcact cagtggcatg aacgtctgtc gggccttgaa
     3121 gcgattcggc aagactatgc gacaggcgat ctggccgtac gcattgccag cctcgacaag
     3181 ctgaaccggc tgcttggcga agccggcgtg gatatttcgc gtgagagcgg cgccatgtat
     3241 gtcggtcgct acccggtcta cgaggattgt tcgcgcaata tcgatatcag cctgggtggc
     3301 gccttacttg atcaggtcaa tgccgatctg gcgccgttga tgcgcattca tcaatggctg
     3361 atcaaagcat gcgctcggca actgcatggt ttttacgctc aggtatggca acgcctgcag
     3421 gcagaggacg agcataatcc ggtgaacttc ctggcctttc ttggcgctgt gcaaccagta
     3481 attgcacacg ccgaggcggg catcatcgaa gaactggacg cagtgctggc agacgcctgg
     3541 cagcaggtcc tgagcgataa acacgaccct gaacaggtgc agttgaccca tgaggacata
     3601 gagcggctga ttgtcgagct gaacacgcgt ctggatgttc ggacgttcag cgtatttggc
     3661 agccactttc actctccgga tttcctgatt tccagtacct cgacagacgc gctgaaccag
     3721 ggcgactact caatcattct gggagaggtc catccgggcg tgcacaccct cagtcagccg
     3781 gtagccgagc ctttcggacc gttcaataaa gagatcgagc aggaggttcg gcaaatattc
     3841 gacggacctc gtctggtgct tgccgactcg ccggacagtt atcagcgttc gcatatcgat
     3901 tggcctttgc tcgagtgcta ccaacaactg atactgcctt ctggcggcgg ttgcgtgccg
     3961 gcggatcagc gctttgccgt cggccgtgct cgtcttgtca tgaacgaggg acgtttgcgc
     4021 gtcgaggata ttgccggcca attcagcgaa gacttggtgt gcgtgtatcc cactcctatg
     4081 caccgactgg gttttgcctt ggctggcagc gtcgtggcca agaacgaccg acgccgaata
     4141 ggcttgggca agaccttgta caaacgcgcc tcgtggtggt tcagtccaga gcaactgccg
     4201 tgctctgaat tcagtgtgga caagctcgac gacgtgctgg catggcgagc gtgggccgtc
     4261 gagcacggtt tgccccgtta tgtgttcgcc aagatcgata tcgaacccaa gccgattttc
     4321 attgatttcg ataacccgtt gtcactcgac ggtgtcagta attcgatgaa gaaagcgggg
     4381 cacgtcaagt tttccgagat gtgtcctgct ccggaccaac tgtggctcga agaaacgcgc
     4441 gggcatttct gttgtgaaat acgtacaacc tttagggaca atggagtaac gcgtgatgaa
     4501 taagcggccg cataatgctt aagtcgaaca gaaagtaatc gtattgtaca cggccgcata
     4561 atcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt ccccatctta
     4621 gtatattagt taagtataag aaggagatat acatatggca gatctcaatt ggatatcggc
     4681 cggccacgcg atcgctgacg tcggtaccct cgagtctggt aaagaaaccg ctgctgcgaa
     4741 atttgaacgc cagcacatgg actcgtctac tagcgcagct taattaacct aggctgctgc
     4801 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     4861 tttgctgaaa cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa
     4921 accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac
     4981 aagctgacga ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta
     5041 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     5101 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     5161 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     5221 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     5281 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     5341 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     5401 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     5461 gcgagacgaa atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     5521 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     5581 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     5641 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     5701 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     5761 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     5821 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     5881 caagacgttt cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt
     5941 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6001 ataggcatgc agcgctcttc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     6061 ctgcggcgag cggtgtcagc tcactcaaaa gcggtaatac ggttatccac agaatcaggg
     6121 gataaagccg gaaagaacat gtgagcaaaa agcaaagcac cggaagaagc caacgccgca
     6181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagcca
     6241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6361 gggaagcgtg gcgctttctc atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt
     6421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6541 cattggtaac tgatttagag gactttgtct tgaagttatg cacctgttaa ggctaaactg
     6601 aaagaacaga ttttggtgag tgcggtcctc caacccactt accttggttc aaagagttgg
     6661 tagctcagcg aaccttgaga aaaccaccgt tggtagcggt ggtttttctt tatttatgag
     6721 atgatgaatc aatcggtcta tcaagtcaac gaacagctat tccgttactc tagatttcag
     6781 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     6841 tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     6901 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     6961 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7021 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     7081 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     7141 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     7201 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     7261 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     7321 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     7381 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     7441 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     7501 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     7561 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     7621 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     7681 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     7741 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     7801 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     7861 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     7921 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     7981 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     8041 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     8101 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     8161 aggaaattaa tacgactcac tatagggga
//
