LOCUS       sequence-305921-        8367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TglC, TglD, TglE, TglF in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8367)
  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 8367)
  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: pACYCDuet1-tglCDEF
FEATURES             Location/Qualifiers
     source          1..8367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        74..92
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    93..117
                     /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             173..190
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        304..322
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    323..347
                     /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             4510..4521
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4816..4860
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(4898..4916)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4912..4959
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5180..5839)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5754..5773
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5840..5942)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6331..6348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6465..7010)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(6545..6564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7235..8317)
                     /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     8279..8298
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(8318..8367,1..28))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
       61 tgcattagga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
      121 gtagaaataa ttttgtttaa ctttaataag gagatatacc atgggcagca gccatcacca
      181 tcatcaccac agccaggatc cgaattcgag ctcggcgcgc ctgcaggtcg acaagcttgc
      241 ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc gcataatcga
      301 aattaatacg actcactata ggggaattgt gagcggataa caattcccca tcttagtata
      361 ttagttaagt ataagaagga gatatacata tgtcgtgcat tttggatgat gagcgctgta
      421 tccccgagtt gttgacgcag cttcgcagtt tgtcgctcga ctttctgtct ggcgcacaca
      481 ccggcgctgc ggtcgacacg cgctctgatg tcctgactca gcaagcggaa atgcccgagg
      541 aggggctcgg ttgcctggat gctctgagga cttattggca gcgctacgct gatgggcaca
      601 gtcgcagcac aggccctcgg tactatggct tcgtgacagg cggggtgacg cccgctgcac
      661 ttgccggaga ctggctggtt tccgtccttg accagaacgt cgccactgag cgccattcga
      721 tcgctgcatt catcgaagct caagtgctga cgttcatcag taacctgctg aagcttccgg
      781 cgggtttgtt tcagggtgtt ttgacgaccg gagcgactgc ggcaaatctg gtggggctgt
      841 ccagcgctcg tgagtggtgc ggtgagcagg cgggagtcag cattgctcac gaaggaattg
      901 cagcagcccc tccgatagcg gtattcgggg cgtctgccca ctcaagtatt atcaaaagtc
      961 tgggtgtgct gggcatcggg cgcaaaaata tagtgtcaat acgcagtttg cctgatcgtg
     1021 aagccatgga tattcaacac ctggaagaag cgctgaaaag cagtgtttct acttctcgaa
     1081 ttgtcgtggc cagcgccggt accgtgaata gcggtgattt cgacgacctc aatggtattg
     1141 ctgatttatg catgaggtac aacgcgtggt tgcatgtcga tgcggcattc ggtgcctttg
     1201 ctcgttgttc tcctcgttac gaacacttga tcgaggggct ggagcgtgct gattcgataa
     1261 cggcggatgc tcacaagtgg cttaacgttc cctatgactg tggaattgcc ctgacccgac
     1321 atacggcgtt tcatgagcgg agcttttccg ctgacgcgcc ctatgtgccc atggggggcg
     1381 agcttcctgc tttcatgaac cgaggtgttg aacagtcgcg tcgctatcgt gcgttgccgt
     1441 tatggatgac gcttatggct tacggcaagt cgggttaccg cgagttggta gagcgtaatt
     1501 gtgatgttgt acgccagatc gcttcgtgga ttgacgggtg tgaatactac gacgtgttgt
     1561 gccctgtcta tttgaacatc gtgatgtttc gctgccgacg tgtcggcggg cgtgatcatg
     1621 acgcgcacag cgtggcagtg ctggaaagaa tcaatgccac aggtgcgctg tttcttacgc
     1681 ccagtacgta caacggggaa aaggtatttc gtctggctgt ctgtaactgg atgaccacac
     1741 gcagtgatgt cgcaattgtg atcaatgcct tacaccacgc ggtggctggt tattgagagc
     1801 agcactgtta tcaaacgata aggaatgtca tacatgtctt cgcaagtgcc tgaaaatacc
     1861 ccttcgacca aggggctgtt cggcctgttt tgcctggcca gctatctgct gtccctgtct
     1921 tacggcacca cttttctact ggccctgatg ctgggctctc atggcggaag cgagtctgac
     1981 gcaggcacag tgatttcagt ggccatgctg agcacatttc tcgccgtcat tttttcaggg
     2041 cacctgaccg atatcatcgg tgcgccccgg gctatcgctg ccggaggtct gacgttgatg
     2101 attgcatgca tcgggtttgc catgaccccg acgtatggcc cggcattatt gatattcggt
     2161 ctgatactgg gtttcggttg gggtgtgttc tacacactgg ggccgatcat tgttgcaatg
     2221 atcatcgagc ctgcgcgtcg ggtgaagtat ttcgctttgc tgtccggcag catgatgagt
     2281 gggatcggta ccgggccttt ggcagggcgc ggtgcagtgg caatggggta tccggttgaa
     2341 tcggcgttcg tcgtcgcggc cgctgccagc ctgctgggtg cagtcgtgtt tttcctgata
     2401 gctccgaaaa tcaggctgca acaggctgtt ctcgggcctg tggccgtgtg ccgcatcact
     2461 acaggtgctg ccgctcgtgt actgcgctcc aaggcgttct ttcctatcgt gatggtggga
     2521 ttgggcggcg caatcttcgg cgggctttcc agctttcaga cctcctacgc cgcattgcat
     2581 catctggact attcgctgtt tttcctcgga ttcatgtgtg cggccatctc ttgccgattg
     2641 ttgattgccg ggttcatcgt caagcgtgat gcttacctgt cttcctgcgt actgacatcc
     2701 ctgatgattg tttcgatact gatgtttgtg ttctcggttg agtcgccatt cagttatttg
     2761 ttggccgcga tcattcttgg cgtgggttat ggcttgacgt attcggtgat caacggcttg
     2821 gcggctaatg aagcgcctaa cggactcacc gcacagtcct tgttgttatt cagtttgtct
     2881 tactttgtag gcgtgttcgg ttttccactg ttggccggaa aaataatcgt ggcttacggc
     2941 atgccctcac tgctgtatac gattctctcg gtagcccttt taaacgggtg tattgcgttg
     3001 ggccgtcttg cctggcgcaa ggtggctgtc gccgccatag cctgaccgta aacgtttcag
     3061 tcaaactgca tcgcgtgacg cgctgcagtt ttcggtgttc ctgtgcaagg taagcaagga
     3121 gattcattgt gcctgttaat gcgcctgaag taataagtct tgaagtggtc ggggatggac
     3181 tggcttcaaa taacgcagcc tggagttttg gcggtgatac gcccaaacat ttcgactcgc
     3241 atgtttcgaa atcggttccc aagtacaagg acggtcacga tatcgttctt tccttgagcg
     3301 acttctttat caaggctgac agcgtctgtt atgaacttgg aagttcgacc ggcacattga
     3361 cacggcaatt ggcagaacgt cacgggcgtt caggcaaatg ggtgggtatc gatgtcgaag
     3421 aggccatgac tcaacaagcc aatcaatcac tcggctgtgc gcccatcagt aacgtcacct
     3481 ttgtgaccga cgacatcctt acctttccgt atgagaagag cgatttcatc gtcgcgtatt
     3541 atacggttca gtttgtccat ccccgcgttc gccaggagct gttcaatcgt atttacgagt
     3601 ctctgaactg ggggggtgca tttgtgctgt tcgagaaagt cagggggccg gatgctcggt
     3661 ttcaggatgt gatcagcagt ttgtatgtcg actacaagat gcagcagggc tatacgccga
     3721 gcgagatcct ggccaagacg cgtagtctta aaggtgtgct ggaaccgttc tccacggccg
     3781 gcaatcagga catgctcgcc cgagccggtt tcaaggacgt gatgagcatt ttcaaataca
     3841 tttgttttga aggtttcttc tgcatcaagt aacagggacg ttgcgatgaa cgattgggct
     3901 atcagccatt acctggatgt gttgcacatg gacagcggtt atcaagccgt tgcactggac
     3961 tgcgaactct gcggcagtga acaccatgag gtgttgctgc cacgtatctg cggtcctgaa
     4021 ccccacggcg ttgatttgcc tgtgttgggg tgtcaggcgt gcggccatat ttttcaaaaa
     4081 tatcgttttg atgcgcagtt ttacaaagac tactacgaca agttttatcg cttgaaactg
     4141 ttcggcaaca gcgcgccgga aaaagcgttt tttctcgacc agattcgtcg tggagaatac
     4201 ctgcgtcaga gcctgcgcga atacctgcca gagagtggtc gcctgctgga tgtcggatgt
     4261 tcagccggtg gcctgatgat cccttttgcc aagaggggct ggtctgtcac cggcaatgac
     4321 cctgatcgcg cttatgtcga atacggaaaa cagatcggcc tgaatatcga gctgacagcg
     4381 gccgaagaaa tggacgccgc aggcgatgtc gatctgatta tcatcaatgg ctcgctggaa
     4441 catatgtacg acgtgcattg cgtcatgcag cgctgtcggc aggcgtcgtc ggaaaacggt
     4501 ctgttattga ttgaaggccg cgccctgggt cacggcctgg agcagggcat gctgacccac
     4561 aatcatcggc gttatctcag cggcaccagc atcgaactgc tgatgcgcat gcacggctgg
     4621 gagccgctgc tgatcaccga gcaaccgtta tgcgggccta ctcgacctgg cgcggttttc
     4681 gtccttggtc gatgcgcagc cccgtcggat gatgtgcagc tcgaacgtgt caaggcgcaa
     4741 gggcgtgagc gcctggacag ttactaccgg gagcgcttcc gggcgctggt gttgccatca
     4801 tgactcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg
     4861 tctactagcg cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa
     4921 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag
     4981 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
     5041 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc
     5101 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
     5161 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     5221 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     5281 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catagtgaaa acgggggcga
     5341 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     5401 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     5461 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     5521 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     5581 tatcccatat caccagctca ccgtctttca ttgccatacg gaactccgga tgagcattca
     5641 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     5701 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     5761 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     5821 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     5881 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     5941 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag
     6001 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt
     6061 cggatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct
     6121 tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa
     6181 agcaccgccg gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg
     6241 agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca
     6301 gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     6361 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     6421 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     6481 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     6541 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
     6601 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
     6661 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
     6721 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
     6781 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
     6841 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
     6901 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
     6961 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
     7021 ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac
     7081 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
     7141 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     7201 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7261 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7321 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     7381 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     7441 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     7501 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     7561 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     7621 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     7681 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     7741 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     7801 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     7861 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     7921 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     7981 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     8041 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     8101 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     8161 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     8221 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     8281 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     8341 cttccgggcg ctatcatgcc ataccgc
//
