LOCUS       sequence-203067-        4696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated codon-opt S. cerevisiae UBP1 and E. coli ClpS in
            an artificial operon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4696)
  AUTHORS   Kunjapur AM, Stork DA, Kuru E, Vargas-Rodriguez O, Landon M, Soll D,
            Church GM
  TITLE     Engineering posttranslational proofreading to discriminate 
            nonstandard amino acids.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Jan 16;115(3):619-624. doi: 
            10.1073/pnas.1715137115. Epub 2018 Jan 4.
  PUBMED    29301968
REFERENCE   2  (bases 1 to 4696)
  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: 
            "10.1073/pnas.1715137115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-01-16- 16"; volume: "115"; issue: "3"; pages: "619-624"
COMMENT     Sequence Label: pZE21/UBP1/ClpS
FEATURES             Location/Qualifiers
     source          1..4696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             83..94
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2633..2719
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2884..3472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2964..2983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3560..3654)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3685..4479)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRQ
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3796..3815)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4406..4425
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4658..4676
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tccctatcag tgatagagat actgagcaca tcagcagtag agatactgag cacatcagca
       61 ggacgcactg accgaattca ttaaagagga gaaaggtacc atggggagtg ggtctttcat
      121 tgctgggctt gtcaacgatg gtaatacgtg ttttatgaac tcggttcttc agtcccttgc
      181 tagtagccgt gaacttatgg agtttttgga taataatgta atccgtacat atgaagaaat
      241 tgaacagaac gagcacaatg aggaaggtaa tggccaagag agcgcacaag atgaggcaac
      301 tcacaaaaaa aacactcgca agggaggtaa ggtctatggg aagcataaaa agaaattaaa
      361 ccgcaaatct tctagcaagg aagacgaaga aaagtcgcaa gaaccagaca ttacgttttc
      421 ggtggcgttg cgtgatctgc tgagcgcatt aaatgctaag tattatcgcg acaaacccta
      481 ctttaagact aactctttat taaaagcgat gagcaagtcc ccgcgcaaaa atatcttgct
      541 tgggtacgat caagaagacg ctcaggaatt ttttcaaaac attcttgcgg agttagaatc
      601 taatgtcaag tcgttaaaca cagaaaagct tgatactaca ccggtagcca agtccgaact
      661 tccagacgat gctctggttg gccaattaaa ccttggtgag gtaggcacag tgtacattcc
      721 cacagaacaa attgacccca attcgatttt acatgacaaa tcgattcaaa actttacccc
      781 ctttaaactg atgaccccgt tggatgggat cacggctgag cgcatcggct gcctgcaatg
      841 cggagagaac gggggaattc gctacagtgt tttcagcgga ttaagtttga acctgccgaa
      901 tgaaaatatt ggaagcactc ttaaactgtc ccagttactg tccgattggt cgaaacccga
      961 gattatcgag ggtgttgaat gcaaccgttg cgctttaaca gctgcgcact cacacttgtt
     1021 tggccaatta aaggagtttg agaagaaacc tgaaggctcg attcccgaaa aacttattaa
     1081 tgccgtaaag gaccgcgtgc accagatcga agaggtcttg gcaaagccgg ttatcgacga
     1141 tgaagattat aaaaaattgc atactgcgaa tatggtccgc aagtgttcaa aaagtaaaca
     1201 aattcttatc tctcgtccac cacctttgtt gtctattcat atcaaccgct ctgttttcga
     1261 cccgcgcacc tacatgattc gcaagaacaa ctccaaggtt ttgttcaagt cacgcttgaa
     1321 cctggcaccc tggtgctgtg atatcaacga aatcaatctt gacgcacgcc ttccgatgtc
     1381 gaagaaggaa aaagcagctc aacaagattc ttctgaagac gagaacattg gcggagagta
     1441 ctatactaaa ttgcatgaac gttttgagca ggagtttgaa gattctgaag aagagaagga
     1501 atacgatgat gcagagggta attatgcatc gcattataac cataccaagg acatctccaa
     1561 ctacgatcca ttgaatggag aagtcgacgg tgtgacttcc gatgatgagg atgaatacat
     1621 tgaagagaca gacgcgttgg ggaataccat caaaaaacgt attattgaac actccgacgt
     1681 ggagaacgaa aacgtgaagg ataatgaaga acttcaggag atcgataacg ttagcttgga
     1741 tgagccaaaa attaatgtcg aggaccagct tgaaacgagt tctgatgagg aagacgttat
     1801 tcctgctcca cccatcaact acgctcgcag ctttagtacg gtcccagcga cccctttaac
     1861 ttactctttg cgcagcgtca tcgtgcacta tgggactcac aactacggac attatattgc
     1921 atttcgcaag tatcgtggat gttggtggcg catctccgat gagacggtct atgtggtaga
     1981 tgaggccgaa gtactgtcaa caccgggggt atttatgctt ttctacgagt atgatttcga
     2041 cgaggagacc ggaaaaatga aagacgactt agaagctatc cagagcaata atgaggaaga
     2101 tgacgagaaa gaacaggaac agaagggtgt ccaggagcca aaagaatccc aggagcaagg
     2161 cgaaggcgaa gaacaagaag aagggcaaga gcaaatgaaa tttgagcgta cggaggatca
     2221 tcgcgacatt tcagggaagg atgtgaatta ataataaaag gagatatacc atgggtaaaa
     2281 cgaacgactg gctggacttt gatcaactgg cggaagaaaa agttcgcgac gcgctaaaac
     2341 cgccatctat gtataaagtg atattagtca atgatgatta cactccgatg gagtttgtta
     2401 ttgacgtgtt acaaaaattc ttttcttatg atgtagaacg tgcaacgcaa ttgatgctcg
     2461 ctgttcacta ccaggggaag gccatttgcg gagtctttac cgccgaggtt gcagaaacca
     2521 aagtggcgat ggtgaacaag tacgcgaggg agaatgagca tccattgctg tgtacgctag
     2581 aaaaagccta aaagcttgat gggggatccc atggtacgcg tgctagaggc atcaaataaa
     2641 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2701 tctcctgagt aggacaaatc cgccgcccta gacctagggc tttcggctgc ggcgagcggt
     2761 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2821 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2881 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2941 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3001 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3061 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3121 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3181 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3241 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3301 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     3361 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3421 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3481 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3541 ggtcatgact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     3601 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     3661 agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     3721 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     3781 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     3841 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     3901 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     3961 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     4021 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     4081 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     4141 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     4201 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     4261 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     4321 caggtcggtc ttgacaaaaa gaacctggcg cccctgcgct gacagccgga acacggcggc
     4381 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     4441 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     4501 ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     4561 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccgacgt
     4621 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4681 tcgtcttcac ctcgag
//
