LOCUS       sequence-305744-        6426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His6-TglBin E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6426)
  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 6426)
  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-tglB
FEATURES             Location/Qualifiers
     source          1..6426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             78..95
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2625..2643
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2644..2668
                     /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             2777..2821
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2859..2877)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2873..2920
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3141..3800)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3715..3734
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3801..3903)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4295..4312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4429..4974)
                     /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(4509..4528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5199..6281)
                     /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     6243..6262
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6282..6359)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        6405..6423
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(6424..6426,1..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 accacagcaa ggatccgatg gaaagctcac
      121 actatttctg gcttcgctcg accgggtttg cggtgcatca cctgacccgc ttggggaaaa
      181 tggccgaatt gccgttgctg aaggattttg aaaccgatta tcgctccctg aatactctac
      241 gcgacagcct gttagagaag tccataagcc actcgtctca ggcctgtcgc aagctgatcc
      301 gcaagctgaa tgaaaacctg ccgctgcaag tgtctgatct tccagaagcg ctgcgtgatg
      361 acgcagtgga tgaattggcg cactggaacg agcggcttgg ccggctcaag caacgtgtcg
      421 aagtggatca ggaatatgcc gtgtttctcg aaagcgcgcg gcaggccctg attgatttcg
      481 tgaacgatga agacgttgag caggccgtgt tcatttccaa cccgaccgcg ctgacccggc
      541 ttcgggagct caggcaagag cgccatgcga gaactgacag ccgcaagaag cagaaactgc
      601 gcctggcctg gagctacgca cagcgattct gttcaaaaaa tgatacgtcg agttttttcg
      661 gcccgctggc ctgggggcgg tttgatcgta cgcaggtcga gcatgttcgt atcacgcagc
      721 acggacccgg ctggatcaga gaacgccata cctttttcga gagctgggtt gtacagcggc
      781 tggtcgagca gttgaatcgc cattgcgttg atgtgcagtt catgccgttg caactcaacc
      841 cgggctgttt tttgaatcaa gacacgttgc atctgcccgt caacaaacgt cgtcaagtca
      901 gtgccttgac ggccagagtg ctgggttaca ttcagtgcca gagcgctgtg gcaccgaccc
      961 tgttcgggtt gcaggccgag ttgcaggatg tcagcgccgg gcaattgcgc gacctgatcg
     1021 atcatctggt cgcccagcag attgtacgtc gcggctggca gatttccccc agggaacgaa
     1081 agcctgttca ggtgctgcac acttttctgg ccgacgcgcg cctgtctgat gactttcgca
     1141 ctcagtggca tgaacgtctg tcgggccttg aagcgattcg gcaagactat gcgacaggcg
     1201 atctggccgt acgcattgcc agcctcgaca agctgaaccg gctgcttggc gaagccggcg
     1261 tggatatttc gcgtgagagc ggcgccatgt atgtcggtcg ctacccggtc tacgaggatt
     1321 gttcgcgcaa tatcgatatc agcctgggtg gcgccttact tgatcaggtc aatgccgatc
     1381 tggcgccgtt gatgcgcatt catcaatggc tgatcaaagc atgcgctcgg caactgcatg
     1441 gtttttacgc tcaggtatgg caacgcctgc aggcagagga cgagcataat ccggtgaact
     1501 tcctggcctt tcttggcgct gtgcaaccag taattgcaca cgccgaggcg ggcatcatcg
     1561 aagaactgga cgcagtgctg gcagacgcct ggcagcaggt cctgagcgat aaacacgacc
     1621 ctgaacaggt gcagttgacc catgaggaca tagagcggct gattgtcgag ctgaacacgc
     1681 gtctggatgt tcggacgttc agcgtatttg gcagccactt tcactctccg gatttcctga
     1741 tttccagtac ctcgacagac gcgctgaacc agggcgacta ctcaatcatt ctgggagagg
     1801 tccatccggg cgtgcacacc ctcagtcagc cggtagccga gcctttcgga ccgttcaata
     1861 aagagatcga gcaggaggtt cggcaaatat tcgacggacc tcgtctggtg cttgccgact
     1921 cgccggacag ttatcagcgt tcgcatatcg attggccttt gctcgagtgc taccaacaac
     1981 tgatactgcc ttctggcggc ggttgcgtgc cggcggatca gcgctttgcc gtcggccgtg
     2041 ctcgtcttgt catgaacgag ggacgtttgc gcgtcgagga tattgccggc caattcagcg
     2101 aagacttggt gtgcgtgtat cccactccta tgcaccgact gggttttgcc ttggctggca
     2161 gcgtcgtggc caagaacgac cgacgccgaa taggcttggg caagaccttg tacaaacgcg
     2221 cctcgtggtg gttcagtcca gagcaactgc cgtgctctga attcagtgtg gacaagctcg
     2281 acgacgtgct ggcatggcga gcgtgggccg tcgagcacgg tttgccccgt tatgtgttcg
     2341 ccaagatcga tatcgaaccc aagccgattt tcattgattt cgataacccg ttgtcactcg
     2401 acggtgtcag taattcgatg aagaaagcgg ggcacgtcaa gttttccgag atgtgtcctg
     2461 ctccggacca actgtggctc gaagaaacgc gcgggcattt ctgttgtgaa atacgtacaa
     2521 cctttaggga caatggagta acgcgtgatg aataagcttg cggccgcata atgcttaagt
     2581 cgaacagaaa gtaatcgtat tgtacacggc cgcataatcg aaattaatac gactcactat
     2641 aggggaattg tgagcggata acaattcccc atcttagtat attagttaag tataagaagg
     2701 agatatacat atggcagatc tcaattggat atcggccggc cacgcgatcg ctgacgtcgg
     2761 taccctcgag tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc acatggactc
     2821 gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat aactagcata
     2881 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga
     2941 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
     3001 agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc tttcgaattt
     3061 ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt ttaagggcac
     3121 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     3181 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     3241 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatagtgaa aacgggggcg
     3301 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     3361 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     3421 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     3481 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     3541 ctatcccata tcaccagctc accgtctttc attgccatac ggaactccgg atgagcattc
     3601 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     3661 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     3721 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     3781 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     3841 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     3901 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     3961 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg
     4021 tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt
     4081 ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg
     4141 caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta tgttggcact
     4201 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     4261 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     4321 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     4381 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     4441 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     4501 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     4561 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     4621 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     4681 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     4741 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     4801 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     4861 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     4921 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     4981 catcttatta atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta
     5041 gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg
     5101 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     5161 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5221 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5281 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     5341 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     5401 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     5461 tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc
     5521 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     5581 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     5641 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     5701 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     5761 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     5821 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     5881 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     5941 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     6001 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     6061 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     6121 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     6181 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     6241 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     6301 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     6361 gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac gactcactat
     6421 agggga
//
