LOCUS       sequence-305641-        4151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His6-AmmA(∆50-56) in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4151)
  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 4151)
  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(∆50-56)
FEATURES             Location/Qualifiers
     source          1..4151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..54
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    55..79
                     /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             135..152
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        407..425
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    426..450
                     /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             559..603
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(641..659)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      655..702
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(923..1582)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1497..1516
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1583..1685)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2077..2094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2211..2756)
                     /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(2291..2310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2981..4063)
                     /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     4025..4044
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4064..4141)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta taggggaatt
       61 gtgagcggat aacaattccc ctgtagaaat aattttgttt aactttaata aggagatata
      121 ccatgggcag cagccatcac catcatcacc acagccagga tccgatgtcc gaaacacagg
      181 ttacagagac agacaatcca gcggaggagc cagccgagat agctgccgaa tcggacgatt
      241 tagcggattt ggacgatatt gagtttgatc tggacgaggt ggagtctaaa atcgcacccc
      301 tggcacttgc ctgactgaaa gagtcccggt cctggtgata agcggccgca taatgcttaa
      361 gtcgaacaga aagtaatcgt attgtacacg gccgcataat cgaaattaat acgactcact
      421 ataggggaat tgtgagcgga taacaattcc ccatcttagt atattagtta agtataagaa
      481 ggagatatac atatggcaga tctcaattgg atatcggccg gccacgcgat cgctgacgtc
      541 ggtaccctcg agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac
      601 tcgtctacta gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca
      661 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt
      721 gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca
      781 taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat
      841 ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc
      901 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta
      961 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     1021 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatagtg aaaacggggg
     1081 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     1141 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     1201 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     1261 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     1321 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc ggatgagcat
     1381 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     1441 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     1501 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     1561 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     1621 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     1681 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac
     1741 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg
     1801 cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca
     1861 gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac
     1921 ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca
     1981 ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg
     2041 tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc
     2101 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     2161 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     2221 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
     2281 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
     2341 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
     2401 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
     2461 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
     2521 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga
     2581 agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa
     2641 gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca
     2701 aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag
     2761 atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg
     2821 tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg
     2881 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     2941 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3001 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3061 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     3121 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     3181 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     3241 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
     3301 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     3361 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     3421 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     3481 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     3541 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     3601 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     3661 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     3721 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     3781 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     3841 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     3901 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     3961 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     4021 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     4081 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     4141 ccgggatctc g
//
