LOCUS       sequence-430198-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid circuit displaying hsCRBN-W384A,Y386A bait (P22 anchor) and 
            driving expression of gIII-neg.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  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 5406)
  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: pSD025A
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(25..52)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(144..230)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     383..400
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(453..1268)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(453..1268)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1176..1195
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1260..1279
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(1409..1954)
                     /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(1489..1508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2246..2263
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4705..4728)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 cctcagcgag tttgtagaaa cgcaaaaagg acatccgtca ggatggcctt ctgcttaatt
       61 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcgca
      121 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
      181 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca
      241 gttccctact ctcgcatggg gagaccccac actaccatcg gcgctacggc gtttcacttc
      301 tgagttcggc atggggtcag gtgggaccac cgcgctactg ccgccaggca aattctgttt
      361 tatcagaccg cttctgcgtt ctgatttaat ctgtatcagg ctgaaaatct tctctcatcc
      421 gccaaaacag ccatgcccat ggcgtacgga tcttagaaaa actcatcgag catcaaatga
      481 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
      541 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
      601 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
      661 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
      721 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
      781 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
      841 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
      901 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
      961 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     1021 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     1081 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     1141 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     1201 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     1261 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgaa
     1321 gagaaaccta cggatcgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     1381 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     1441 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     1501 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     1561 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     1621 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     1681 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     1741 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     1801 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     1861 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     1921 gcaagatatt acgcgcagac caaaacgatc tcaagaagat catcttatta aggagcccca
     1981 tacgatatac gagaggacat ccgggcaaat gcaacgaaag gctcagtcga aagactgggc
     2041 ctttcgttgc gctgcggaga cttagggacc ctaatattta agacttctta atttcaaggc
     2101 accccgggct ttactcgtaa agcttccggc gcgtatgttg tgtcgaccga cctgcaggtg
     2161 cagtaaggag gaaaaaaaaa tgaaaaaatt attattcgca attcctttag ttgttccttt
     2221 ctattctcac tccgctgaaa ctgttcatca ccatcaccat cacgctgaaa ctgttgaaag
     2281 ttgtttagca aaaccccata cagaaaattc atttactaac gtctggaaag acgacaaaac
     2341 tttagatcgt tacgctaact atgagggctg tctgtggaat gctacaggcg ttgtagtttg
     2401 tactggtgac gaaactcagt gttacggtac atgggttcct attgggcttg ctatccctga
     2461 aaatgagggt ggtggctctg agggtggcgg ttctgagggt ggcggttctg agggtggcgg
     2521 tactaaacct cctgagtacg gtgatacacc tattccgggc tatacttata tcaaccctct
     2581 cgacggcact tatccgcctg gtactgagca aaaccccgct aatcctaatc cttctcttga
     2641 ggagtctcag cctcttaata ctttcatgtt tcagaataat aggttccgaa ataggcaggg
     2701 ggcattaact gtttatacgg gcactgttac tcaaggcact gaccccgtta aaacttatta
     2761 ccagtacact cctgtatcat caaaagccat gtatgacgct tactggaacg gtaaattcag
     2821 agactgcgct ttccattctg gctttaatga ggatccattc gtttgtgaat atcaaggcca
     2881 atcgtctgac ctgcctcaac ctcctgtcaa tgctggcggc ggctctggtg gtggttctgg
     2941 tggcggctct gagggtggtg gctctgaggg tggcggttct gagggtggcg gctctgaggg
     3001 aggcggttcc ggtggtggct cttcccaaat ggctcaagtc ggtgacggtg ataattcacc
     3061 tttaatgaat aatttccgtc aatatttacc ttccctccct caatcggttg aatgtcgccc
     3121 ttttgtcttt ggcgctggta aaccttacga gttcagtatc gactgcgata agatcaacct
     3181 gttccgcggt gtctttgcgt ttcttttata tgttgccacc tttatgtatg tattttctac
     3241 gtttgctaac atactgcgta ataaggagtc ttaaacggag ccaatgtacg caaaaaaccc
     3301 cgcttcagcg gggttttttc gccaaaaaaa accccgccct gtcaggggcg gggttttttt
     3361 ttagggccct actgactgtt acaagcaaag aattaccttg tctggggaaa tttcgtcttc
     3421 agtgtcaggg atagtgggca gcagagcaga acgcgtcagg ccccaaaact tctggggaga
     3481 catgtctttc ttagtagccg tgaacttcca gccgatgtga gatgcgcaaa ttttgcattg
     3541 tgcaacagtt gcggctgccc cagggaacca agagtgctcg gtagaaggac gaccgataag
     3601 attaagattg catgctttat ataccgtcag cgtttcgtgg acgtagccat gaggattaac
     3661 gtacgcagcc atcggcccgc aaagagacag actaaagatc tcattcttcg ttgtaatttc
     3721 cgtttcctgg cattgcttac agcacagact agtgcactta ttcataatgt caagttcaca
     3781 acgcaagcgt tggatagccg aaccaatttt caaaagttgg atacgtaaga catcatcaat
     3841 gggaaggcac gcagccacgc ggtacgagaa gtcaattggg ttggaaggaa gggagtcgtc
     3901 ttttaagttt tcgtcccact cgcgtaattg ttttttaata cggtccataa gcgtctctgc
     3961 gtcgtacaac gaatataacc aacgcggcca gcttgtaagg ttggcacaat gaaatttacg
     4021 cttctggtac ttttgccacc acttgtagct acattggtcc tcacgggaca ctggcttcga
     4081 cggaaagatt tgacatttat ttaaggactc caattgcact gctgacatag tagagggcag
     4141 gacgcactct ggtaagattt ggaccttggc ctgctggatt ccatcgcttt gggtacgcaa
     4201 ttccagaact ttgaagcgtt gacgtccgat ggcttttact tttacaattt cgatcccgaa
     4261 gtcctgttct tcacgatatg cgtaaatctc ggctgtggtt ccaaattgtg cttcgcgttc
     4321 ctggacattg ctgtaggcta aaactgcaaa ggtgcggtcc ttctggatca aattacgtac
     4381 catcgaaact tcttgcgggt gaaaaagttg caaaggaagt gtctggcccg gaatcaggat
     4441 catcatcact tgaggcagta cggggataac ctggcaagag tcatcgtcgt gaagcgtacg
     4501 gccgtgaaac tcctccatat cagcccccag atatgtgtga cttgtaggca ggctggtatc
     4561 gaaattgata atgttaggct tttttgcttc tttcgaatct tgatcctcga cttccatctc
     4621 atcctcctcc tcactctctg ctggcagcaa cggcagatgg ttccccatat tatgcgctgc
     4681 gtcctgttga tccccctctc cggccttgtc atcgtcgtcc ttgtagtcga catttgtctg
     4741 tgacaaatcg ccttttaaca ggtagtcggg cgaacactgt aaggctttcg aaagggccaa
     4801 taagttctcc ccattaggct ctgtttcact gcgctcccac tgggaaatgg caacgttact
     4861 cacacctacc atcttcccca gagccgcttg acggatcttt aattttttgc gacgtgcgcg
     4921 aatgcgttct cccatcagtt gagtgtttgc gcttacccag gagatcaatg ggtaacttcc
     4981 acgtggttcg tggcgcgagt gataagcaac attcgtctgg gataaatcgc ccttaagtaa
     5041 atagtcaggc gagcattgca gggccttgct aagcgccaga aggttttcgc cgttcggctc
     5101 cgtttcgcta cgctcccatt gagaaatggc tacgttagat acaccgacca ttttccctaa
     5161 ggccgcctga cggatcttaa gcttcttacg gcgagcgcgg atgcgctcac ccattaactg
     5221 ggtgttcatt tttttttcct ccttactcta gtaaaagtta aacaaaatta tttgtagagg
     5281 gaaaccgttg tggtctccct gaatatagat accgagcctt atgcatgccc gtaaagttat
     5341 ccagcaacca ctcatagacc tagggcagca gatagggacg acgtggtgtt agctgtgctc
     5401 tagaat
//
