LOCUS       sequence-433621-        6781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid circuit displaying hsCRBN bait and driving expression of 
            gIII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6781)
  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 6781)
  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: pJM186A
FEATURES             Location/Qualifiers
     source          1..6781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..951)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      999..1221
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(4593..4616)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(5289..5316)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5408..5494)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5647..5664
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(5710..6501,6502..6570))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6333..6352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6571..6675)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6743..6761
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
       61 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
      121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      181 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
      661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
      721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
      781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
      841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
      901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
      961 ttggtctagg taattttaat cactatacca attgagatgg gctagtcaat gataattact
     1021 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     1681 ggcttacccg tcgtacggtc aagaggacat ccgggcaaat gcaacgaaag gctcagtcga
     1741 aagactgggc ctttcgttgc gctgcggaga cttagggacc ctaatacaag aaagtttgtt
     1801 cattaggcac cccgggcttt actcgtaaag cttccggcgc gtatgttgtg tcgaccgacc
     1861 tgcaggtgca gtaaggagga aaaaaaaatg aaaaaattat tattcgcaat tcctttagtt
     1921 gttcctttct attctcactc cgctgaaact gttgaaagtt gtttagcaaa accccataca
     1981 gaaaattcat ttactaacgt ctggaaagac gacaaaactt tagatcgtta cgctaactat
     2041 gagggctgtc tgtggaatgc tacaggcgtt gtagtttgta ctggtgacga aactcagtgt
     2101 tacggtacat gggttcctat tgggcttgct atccctgaaa atgagggtgg tggctctgag
     2161 ggtggtggtt ctgagggtgg cggttctgag ggtggcggta ctaaacctcc tgagtacggt
     2221 gatacaccta ttccgggcta tacttatatc aaccctctcg acggcactta tccgcctggt
     2281 actgagcaaa accccgctaa tcctaatcct tctcttgagg agtctcagcc tcttaatact
     2341 ttcatgtttc agaataatag gttccgaaat aggcaggggg cattaactgt ttatacgggc
     2401 actgttactc aaggcactga ccccgttaaa acttattacc agtacactcc tgtatcatca
     2461 aaagccatgt atgacgctta ctggaacggt aaattcagag actgcgcttt ccattctggc
     2521 tttaatgagg atccattcgt ttgtgaatat caaggccaat cgtctgacct gcctcaacct
     2581 cctgtcaatg ctggcggcgg ctctggtggt ggttctggtg gcggctctga gggtggtggc
     2641 tctgagggtg gcggttctga gggtggcggc tctgagggag gcggttccgg tggtggctct
     2701 ggttccggtg attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa
     2761 aatgccgatg aaaacgcgct acagtctgac gctaaaggca aacttgattc tgtcgctact
     2821 gattacggtg ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat
     2881 ggtgctactg gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat
     2941 aattcacctt taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa
     3001 tgtcgccctt ttgtctttgg cgctggtaaa ccttacgagt tcagtatcga ctgcgataag
     3061 atcaacctgt tccgcggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta
     3121 ttttctacgt ttgctaacat actgcgtaat aaggagtctt aaacggagcc aatgtacgca
     3181 aaaaaccccg cttcagcggg gttttttcgc caaaaaaaac cccgccctgt caggggcggg
     3241 gttttttttt agggccctac tgactgttac aagcaaagaa ttaccttgtc tggggaaatt
     3301 tcgtcttcag tgtcagggat agtgggcagc agagcagaac gcgtcaggcc ccaaaacttc
     3361 tggggagaca tgtctttctt agtagccgtg aacttccagc cgatgtgaga tgcgcaaatt
     3421 ttgcattgtg caacagtcca ggcgtaccca gggaaccaag agtgctcggt agaaggacga
     3481 ccgataagat taagattgca tgctttatat accgtcagcg tttcgtggac gtagccatga
     3541 ggattaacgt acgcagccat cggcccgcaa agagacagac taaagatctc attcttcgtt
     3601 gtaatttccg tttcctggca ttgcttacag cacagactag tgcacttatt cataatgtca
     3661 agttcacaac gcaagcgttg gatagccgaa ccaattttca aaagttggat acgtaagaca
     3721 tcatcaatgg gaaggcacgc agccacgcgg tacgagaagt caattgggtt ggaaggaagg
     3781 gagtcgtctt ttaagttttc gtcccactcg cgtaattgtt ttttaatacg gtccataagc
     3841 gtctctgcgt cgtacaacga atataaccaa cgcggccagc ttgtaaggtt ggcacaatga
     3901 aatttacgct tctggtactt ttgccaccac ttgtagctac attggtcctc acgggacact
     3961 ggcttcgacg gaaagatttg acatttattt aaggactcca attgcactgc tgacatagta
     4021 gagggcagga cgcactctgg taagatttgg accttggcct gctggattcc atcgctttgg
     4081 gtacgcaatt ccagaacttt gaagcgttga cgtccgatgg cttttacttt tacaatttcg
     4141 atcccgaagt cctgttcttc acgatatgcg taaatctcgg ctgtggttcc aaattgtgct
     4201 tcgcgttcct ggacattgct gtaggctaaa actgcaaagg tgcggtcctt ctggatcaaa
     4261 ttacgtacca tcgaaacttc ttgcgggtga aaaagttgca aaggaagtgt ctggcccgga
     4321 atcaggatca tcatcacttg aggcagtacg gggataacct ggcaagagtc atcgtcgtga
     4381 agcgtacggc cgtgaaactc ctccatatca gcccccagat atgtgtgact tgtaggcagg
     4441 ctggtatcga aattgataat gttaggcttt tttgcttctt tcgaatcttg atcctcgact
     4501 tccatctcat cctcctcctc actctctgct ggcagcaacg gcagatggtt ccccatatta
     4561 tgcgctgcgt cctgttgatc cccctctccg gccttgtcat cgtcgtcctt gtagtcaaca
     4621 ttcgaatcag aggtgccatt gagcagccag tcaacactta cgccaagagc tgacgcaagt
     4681 tctggtaaaa agcgtggtcg cttagtttta ccgttttcga gctgctctat agactgctgg
     4741 gtagtcccca ccttttgagc aagttcagcc tggttaagtc caagctgaat tcttttgctt
     4801 tttaccctgc tgctgatact agctctaacc atgctaatca atggatattt ccctttgggc
     4861 tcaacgtgcc caacaaatct aacattcgaa tcagaggtgc cattgagcag ccagtcaaca
     4921 cttacgccaa gagctgacgc aagttctggt aaaaagcgtg gtcgcttagt tttaccgttt
     4981 tcgagctgct ctatagactg ctgggtagtc cccacctttt gagcaagttc agcctggtta
     5041 agtccaagct gaattctttt gctttttacc ctgctgctga tactcatttt tttttcctcc
     5101 ttactctagt aaaagttaaa caaaattatt tgtagaggga aaccgttgtg gtctccctga
     5161 atatagatac cgagccttat gcatgcccgt aaagttatcc agcaaccact catagaccta
     5221 gggcagcaga tagggacgac gtggtgttag ctgtgctcta gaatcctcag cgagtttgta
     5281 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
     5341 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
     5401 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     5461 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca
     5521 tggggagacc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg
     5581 tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag accgcttctg
     5641 cgttctgatt taatctgtat caggctgaaa atcttctctc atccgccaaa acagccatgc
     5701 ccatggcgtt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6301 accgagttgc tcttgcccag cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6721 tatcatgaca ttaacctata aaaataggcg tatcacgagg cccttaggcc aaataggccg
     6781 t
//
