LOCUS       sequence-430203-        6789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid circuit displaying mmCRBN bait and driving expression of 
            gIII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6789)
  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 6789)
  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: pSD094B
FEATURES             Location/Qualifiers
     source          1..6789
                     /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"
     protein_bind    4514..4525
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             complement(4601..4624)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(5297..5324)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5416..5502)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5655..5672
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(5718..6509,6510..6578))
                     /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     6341..6360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6579..6683)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6751..6769
                     /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 gtaaaaaagg aaaaaaatga aaaaattatt attcgcaatt cctttagttg
     1921 ttcctttcta ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa ccccatacag
     1981 aaaattcatt tactaacgtc tggaaagacg acaaaacttt agatcgttac gctaactatg
     2041 agggctgtct gtggaatgct acaggcgttg tagtttgtac tggtgacgaa actcagtgtt
     2101 acggtacatg ggttcctatt gggcttgcta tccctgaaaa tgagggtggt ggctctgagg
     2161 gtggtggttc tgagggtggc ggttctgagg gtggcggtac taaacctcct gagtacggtg
     2221 atacacctat tccgggctat acttatatca accctctcga cggcacttat ccgcctggta
     2281 ctgagcaaaa ccccgctaat cctaatcctt ctcttgagga gtctcagcct cttaatactt
     2341 tcatgtttca gaataatagg ttccgaaata ggcagggggc attaactgtt tatacgggca
     2401 ctgttactca aggcactgac cccgttaaaa cttattacca gtacactcct gtatcatcaa
     2461 aagccatgta tgacgcttac tggaacggta aattcagaga ctgcgctttc cattctggct
     2521 ttaatgagga tccattcgtt tgtgaatatc aaggccaatc gtctgacctg cctcaacctc
     2581 ctgtcaatgc tggcggcggc tctggtggtg gttctggtgg cggctctgag ggtggtggct
     2641 ctgagggtgg cggttctgag ggtggcggct ctgagggagg cggttccggt ggtggctctg
     2701 gttccggtga ttttgattat gaaaagatgg caaacgctaa taagggggct atgaccgaaa
     2761 atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa acttgattct gtcgctactg
     2821 attacggtgc tgctatcgat ggtttcattg gtgacgtttc cggccttgct aatggtaatg
     2881 gtgctactgg tgattttgct ggctctaatt cccaaatggc tcaagtcggt gacggtgata
     2941 attcaccttt aatgaataat ttccgtcaat atttaccttc cctccctcaa tcggttgaat
     3001 gtcgcccttt tgtctttggc gctggtaaac cttacgagtt cagtatcgac tgcgataaga
     3061 tcaacctgtt ccgcggtgtc tttgcgtttc ttttatatgt tgccaccttt atgtatgtat
     3121 tttctacgtt tgctaacata ctgcgtaata aggagtctta aacggagcca atgtacgcaa
     3181 aaaaccccgc ttcagcgggg ttttttcgcc aaaaaaaacc ccgccctgtc aggggcgggg
     3241 ttttttttta gggccctact gactgttaaa gacacaggat caccttatcg ggactaattt
     3301 catcctctgt ctctggaatt gtaggtaaaa gcgctgaacg ggttaacccc cagaacttct
     3361 ggggactcat atcctttttt gtggcggtaa atttccaacc aatgtgtgaa gcacagattt
     3421 tgcactgcgc aatggtccat gcgtaaccgg gaaaccaact gtggactgtg ctgggacgac
     3481 caataagatt caagttcgag gccttgtata cagtcagtgt ttcatggacg tagccatgcg
     3541 gattcacata agcagccata ggcccacaca atgacaagct gaagatttca ttcttcgtgg
     3601 taatttcagt ctcttggcat tgcttacagc acaatgacgt acatttattc atgatatcca
     3661 attcacagcg taaacgctga attgccgagc cgatttttaa taattgaata cgtaaaacat
     3721 catcgattgg aagacaagca gcaacgcggt acgagaagtc gataggattc tcaggcaggg
     3781 aatcgtcctt taagttttca tcccactcac gcaactgttt tttgatacga tccatcaaag
     3841 tttcagcgtc gtataacgaa tacaaccagc ggggccaaga agttaaattt gcacagtgga
     3901 atttgcgttt ttggtacttc tgccaccact tgcatgagta ttggtcctcc cacgaaatgg
     3961 gcttagacgg gaagacctgg catttgttca gagactccag ttgaactgca ctcatggtgg
     4021 aaggtaaaac acattctgga aggatttgta ccttggcttg ttgaatcccg tcactctgtg
     4081 tacgtaactc cagaaccttg aaacgttggc gtccgattgc tttgactttg actacctcga
     4141 taccgaactc ctgttcttcg cggtaggcat aaatttctgc tgttgtccca aactgtgctt
     4201 cgcgttcttg gacattgcta taagccaaaa ccgcgaaagt acggtctttc tgaattaaat
     4261 tacgaaccat cgatacttct tgggggtgcg ataattgaag aggaagggtc tgccccggaa
     4321 tcagaatcat aagaacttca ggtaatacgg gaattacttg gcatgaatca tcgtcatgaa
     4381 gcgtgcgccc gtgaaattct tccatatctg cacctaagta ggtatggcta gtgggaagtg
     4441 aggtgtcaaa attaatgatg ttcggtttgc gtgcttcttt ggagtcctga tcctctactt
     4501 ccatttcaat ttcgtcatcc tcatcctcgg aatctgccgg taacaacgga agatgattgc
     4561 ccatattgtg ggctgcatct tgttgatccc cctcccctgc cttgtcatcg tcgtccttgt
     4621 agtcaacatt cgaatcagag gtgccattga gcagccagtc aacacttacg ccaagagctg
     4681 acgcaagttc tggtaaaaag cgtggtcgct tagttttacc gttttcgagc tgctctatag
     4741 actgctgggt agtccccacc ttttgagcaa gttcagcctg gttaagtcca agctgaattc
     4801 ttttgctttt taccctgctg ctgatactag ctctaaccat gctaatcaat ggatatttcc
     4861 ctttgggctc aacgtgccca acaaatctaa cattcgaatc agaggtgcca ttgagcagcc
     4921 agtcaacact tacgccaaga gctgacgcaa gttctggtaa aaagcgtggt cgcttagttt
     4981 taccgttttc gagctgctct atagactgct gggtagtccc caccttttga gcaagttcag
     5041 cctggttaag tccaagctga attcttttgc tttttaccct gctgctgata ctcatttttt
     5101 tttcctcctt actctagtaa aagttaaaca aaattatttg tagagggaaa ccgttgtggt
     5161 ctccctgaat atagataccg agccttatgc atgcccgtaa agttatccag caaccactca
     5221 tagacctagg gcagcagata gggacgacgt ggtgttagct gtgctctaga atcctcagcg
     5281 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttaa tttgatgcct
     5341 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttcg caacgttcaa
     5401 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     5461 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     5521 ctctcgcatg gggagacccc acactaccat cggcgctacg gcgtttcact tctgagttcg
     5581 gcatggggtc aggtgggacc accgcgctac tgccgccagg caaattctgt tttatcagac
     5641 cgcttctgcg ttctgattta atctgtatca ggctgaaaat cttctctcat ccgccaaaac
     5701 agccatgccc atggcgttta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5761 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5821 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5881 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5941 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6001 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6061 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6121 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6181 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6241 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6301 atgcggcgac cgagttgctc ttgcccagcg tcaatacggg ataataccgc gccacatagc
     6361 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6421 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6481 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6541 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6601 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6661 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     6721 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc cttaggccaa
     6781 ataggccgt
//
