LOCUS       sequence-305754-        3954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His6-TglA-Cys in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3954)
  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 3954)
  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)
FEATURES             Location/Qualifiers
     source          1..3954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..41
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    42..66
                     /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             122..139
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        378..396
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    397..421
                     /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             530..574
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(612..630)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      626..673
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(903..1718)
                     /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(903..1718)
                     /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     1626..1645
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1719..1810)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1826..2575)
                     /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(1836..1853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2076..2095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2771..3853)
                     /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     3815..3834
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3854..3931)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tgcgactcct gcattaggaa attaatacga ctcactatag gggaattgtg agcggataac
       61 aattcccctg tagaaataat tttgtttaac tttaataagg agatatacca tgggcagcag
      121 ccatcaccat catcaccaca gccaggatcc gatgggacaa cccaacgtgc agtccgttga
      181 aaaccagcaa gcgtccggcg atgtaaaaga tctcgaaaac actccgcagg ccacagagga
      241 ggcgttgttt gaagagtttg acctggatga cattgaagtc atcgagagca aggtctttgc
      301 ctgctgaagc ttgcggccgc ataatgctta agtcgaacag aaagtaatcg tattgtacac
      361 ggccgcataa tcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
      421 cccatcttag tatattagtt aagtataaga aggagatata catatggcag atctcaattg
      481 gatatcggcc ggccacgcga tcgctgacgt cggtaccctc gagtctggta aagaaaccgc
      541 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
      601 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
      661 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
      721 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
      781 accgacgaca agctgacgac cgggtctccg caagtggcac ttttcgggga aatgtgcgcg
      841 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
      901 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
      961 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1021 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1081 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1141 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1201 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1261 tgcgcctgag cgagacgaaa tacgcggtcg ctgttaaaag gacaattaca aacaggaatc
     1321 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1381 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     1441 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1501 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1561 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     1621 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     1681 ggcctagagc aagacgtttc ccgttgaata tggctcatac tcttcctttt tcaatattat
     1741 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1801 aataaacaaa taggcatgca gcgctcttcc gcttcctcgc tcactgactc gctacgctcg
     1861 gtcgttcgac tgcggcgagc ggtgtcagct cactcaaaag cggtaatacg gttatccaca
     1921 gaatcagggg ataaagccgg aaagaacatg tgagcaaaaa gcaaagcacc ggaagaagcc
     1981 aacgccgcag gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2041 ctcaagccag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2161 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgttggtatc tcagttcggt
     2221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2341 ggcagcagcc attggtaact gatttagagg actttgtctt gaagttatgc acctgttaag
     2401 gctaaactga aagaacagat tttggtgagt gcggtcctcc aacccactta ccttggttca
     2461 aagagttggt agctcagcga accttgagaa aaccaccgtt ggtagcggtg gtttttcttt
     2521 atttatgaga tgatgaatca atcggtctat caagtcaacg aacagctatt ccgttactct
     2581 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     2641 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     2701 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     2761 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2821 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     2881 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     2941 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     3001 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     3061 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     3121 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     3181 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     3241 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     3301 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     3361 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     3421 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     3481 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     3541 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     3601 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     3661 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     3721 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     3781 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     3841 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     3901 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc ctta
//
