LOCUS       sequence-305633-        4148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His6-AmmA in E. Coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4148)
  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 4148)
  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
FEATURES             Location/Qualifiers
     source          1..4148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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             120..137
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        389..407
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    408..432
                     /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             541..585
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(623..641)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      637..684
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(905..1564)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1479..1498
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1565..1667)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2059..2076)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2193..2738)
                     /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(2273..2292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2963..4045)
                     /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     4007..4026
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4046..4123)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgactcctgc attaggaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctgta gaaataattt tgtttaactt taataaggag atataccatg ggcagcagcc
      121 atcaccatca tcaccacagc caggatccga tgtccgaaac acaggttaca gagacagaca
      181 atccagcgga ggagccagcc gagatagctg ccgaatcgga cgatttagcg gatttggacg
      241 atattgagtt tgatctggac gaggtggagt ctaaaatcgc acccctggca cttgccctga
      301 aagagtcccg gtcctggtga taagcggccg cataatgctt aagtcgaaca gaaagtaatc
      361 gtattgtaca cggccgcata atcgaaatta atacgactca ctatagggga attgtgagcg
      421 gataacaatt ccccatctta gtatattagt taagtataag aaggagatat acatatggca
      481 gatctcaatt ggatatcggc cggccacgcg atcgctgacg tcggtaccct cgagtctggt
      541 aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg actcgtctac tagcgcagct
      601 taattaacct aggctgctgc caccgctgag caataactag cataacccct tggggcctct
      661 aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca
      721 ctgcttccgg tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga
      781 ccctgccctg aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct
      841 tattatcact tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa
      901 aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
      961 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     1021 tgtcgccttg cgtataatat ttgcccatag tgaaaacggg ggcgaagaag ttgtccatat
     1081 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     1141 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     1201 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     1261 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     1321 gctcaccgtc tttcattgcc atacggaact ccggatgagc attcatcagg cgggcaagaa
     1381 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     1441 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     1501 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     1561 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     1621 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     1681 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
     1741 cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca
     1801 acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc
     1861 agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga
     1921 catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg
     1981 aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata
     2041 caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg
     2101 agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa
     2161 cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag
     2221 catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac
     2281 caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc
     2341 ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg
     2401 gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg
     2461 cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact
     2521 ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag
     2581 gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca
     2641 aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt
     2701 cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga
     2761 taaaatattt ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
     2821 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
     2881 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     2941 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3001 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3061 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     3121 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     3181 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3241 agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     3301 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     3361 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     3421 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     3481 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     3541 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     3601 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     3661 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     3721 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     3781 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     3841 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     3901 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     3961 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4021 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     4081 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     4141 cccttatg
//
