LOCUS       sequence-430017-        3120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For hybrid circuit activation. Expresses E. coli RNAP omega subunit 
            used to FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3120)
  AUTHORS   Mercer JAM, DeCarlo SJ, Roy Burman SS, Sreekanth V, Nelson AT, 
            Hunkeler M, Chen PJ, Donovan KA, Kokkonda P, Tiwari PK, Shoba VM, 
            Deb A, Choudhary A, Fischer ES, Liu DR
  TITLE     Continuous evolution of compact protein degradation tags regulated 
            by selective molecular glues.
  JOURNAL   Science. 2024 Mar 15;383(6688):eadk4422. doi: 
            10.1126/science.adk4422. Epub 2024 Mar 15.
  PUBMED    38484051
REFERENCE   2  (bases 1 to 3120)
  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"; journalName: "Science."; 
            date: "2024-03-15"; volume: "383(6688):eadk4422. doi"; pages: " 
            10.1126/science.adk4422. Epub 2024 Mar 15"
COMMENT     Sequence Label: pJM166B
FEATURES             Location/Qualifiers
     source          1..3120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..792)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(793..884)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      893..1481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1382..1401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1635..1652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1667..1807
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1733..1752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1893..1915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1909..2100)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             2440..2712
                     /codon_start=1
                     /gene="E. coli rpoZ"
                     /product="DNA-directed RNA polymerase subunit omega"
                     /label=RNAP omega
                     /note="promotes RNA polymerase assembly"
                     /translation="MARVTVQDAVEKIGNRFDLVLVAARRARQMQVGGKDPLVPEENDK
                     TTVIALREIEEGLINNQILDVRERQEQQEQEAAELQAVTAIAEGRR"
     CDS             2722..2751
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
ORIGIN
        1 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
       61 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
      121 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
      181 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
      241 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
      301 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
      361 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
      421 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
      481 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
      541 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
      601 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
      661 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
      721 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
      781 cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
      841 catgagcgga tacatatttg aatgtattta gaaaaatagg ccaaataggc cgttgagatc
      901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      961 tttgtttgcc ggatcaagag ctaccacctc tttttccgaa ggtaactggc ttcagcagag
     1021 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1561 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1621 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     1681 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     1741 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     1801 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     1861 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     1921 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     1981 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     2041 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     2101 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     2161 gaggatgctc acgatacggg ttactgatga tgaacaagag gacatccggt caaataaaac
     2221 gaaaggctca gtcgaaagac tgggcctttc gttttgctga ggattctaga gcacagctaa
     2281 caccacgtcg tccctatctg ctgccctagg tctatgagtg gttgctggat aactttacgg
     2341 gcatgcataa ggctcggtat ctatattcag ggagaccaca acggtttccc tctacaaata
     2401 attttgttta acttttacta gagtaaggag gaaaaaaaaa tggcacgcgt aactgttcag
     2461 gacgctgtag agaaaattgg taaccgtttt gacctggtac tggtcgccgc gcgtcgcgct
     2521 cgtcagatgc aggtaggcgg aaaggatccg ctggtaccgg aagaaaacga taaaaccact
     2581 gtaatcgcgc tgcgcgaaat cgaagaaggt ctgatcaaca accagatcct cgacgttcgc
     2641 gaacgccagg aacagcaaga gcaggaagcc gctgaattac aagccgttac cgctattgct
     2701 gaaggtcgtc gtgcggccgc ggaacaaaag cttatttctg aagaggactt ggggcgtgtt
     2761 gcaatcttgt ggcatgagat gtggcacgaa ggtctggaag aagcctcacg tctttatttt
     2821 ggggaacgca atgttaaagg catgttcgaa gttcttgagc cgttgcacgc tatgatggag
     2881 cgcggccccc aaacccttaa agaaacatca tttaaccagg cgtatggtcg tgacttaatg
     2941 gaagcccaag agtggtgccg caaatacatg aaatcgggaa atgttaagga tttaactcaa
     3001 gcctgggatt tatactatca tgtcttccgc cgcatttcgt aacttaatta acggcactcc
     3061 tcagcaaata taatgaccct cttgataacc caagagggca ttttttaatg cccatggcgt
//
