LOCUS       sequence-305638-        4145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His6-AmmA(∆Trp56) in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4145)
  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 4145)
  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-ammA(∆Trp56)
FEATURES             Location/Qualifiers
     source          1..4145
                     /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        388..406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    407..431
                     /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             540..584
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(622..640)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      636..683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(904..1563)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1478..1497
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1564..1666)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2058..2075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2192..2737)
                     /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(2272..2291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2962..4044)
                     /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     4006..4025
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4045..4122)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tgcgactcct gcattaggaa attaatacga ctcactatag gggaattgtg agcggataac
       61 aattcccctg tagaaataat tttgtttaac tttaataagg agatatacca tgggcagcag
      121 ccatcaccat catcaccaca gccaggatcc gatgtccgaa acacaggtta cagagacaga
      181 caatccagcg gaggagccag ccgagatagc tgccgaatcg gacgatttag cggatttgga
      241 cgatattgag tttgatctgg acgaggtgga gtctaaaatc gcacccctgg cacttgccct
      301 gaaagagtcc cggtcctgat aagcggccgc ataatgctta agtcgaacag aaagtaatcg
      361 tattgtacac ggccgcataa tcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      421 ataacaattc cccatcttag tatattagtt aagtataaga aggagatata catatggcag
      481 atctcaattg gatatcggcc ggccacgcga tcgctgacgt cggtaccctc gagtctggta
      541 aagaaaccgc tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt
      601 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      661 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
      721 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
      781 cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt
      841 attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa
      901 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
      961 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     1021 gtcgccttgc gtataatatt tgcccatagt gaaaacgggg gcgaagaagt tgtccatatt
     1081 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     1141 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     1201 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     1261 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     1321 ctcaccgtct ttcattgcca tacggaactc cggatgagca ttcatcaggc gggcaagaat
     1381 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     1441 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     1501 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     1561 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     1621 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg
     1681 ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc
     1741 gaagtgatct tccgtcacag gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa
     1801 cttactgatt tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca
     1861 gctgtccctc ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac
     1921 atcagcgcta gcggagtgta tactggctta ctatgttggc actgatgagg gtgtcagtga
     1981 agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac
     2041 aggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg actgcggcga
     2101 gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa gatacttaac
     2161 agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc cctgacaagc
     2221 atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc
     2281 aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg
     2341 gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg
     2401 taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc
     2461 gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg
     2521 gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg
     2581 ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa
     2641 agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt tttcgttttc
     2701 agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
     2761 aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     2821 catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg
     2881 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     2941 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3001 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     3061 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     3121 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     3181 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     3241 gatgtccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     3301 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     3361 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     3421 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     3481 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     3541 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     3601 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     3661 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     3721 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     3781 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     3841 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     3901 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     3961 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     4021 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     4081 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     4141 cctta
//
