LOCUS       sequence-430199-        7662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid circuit displaying hsCRBN-W384A,Y386A bait (P22 anchor) and 
            driving expression of luxAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7662)
  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 7662)
  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: pSD139A
FEATURES             Location/Qualifiers
     source          1..7662
                     /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             1890..2969
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     CDS             3033..4013
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="KFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENHF
                     SDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILGF
                     SDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAYT
                     PGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEID
                     HQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECITA
                     AKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHTEYT"
     CDS             complement(5444..5467)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(6170..6197)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6289..6375)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6528..6545
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(6591..7382,7383..7451))
                     /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     7214..7233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7452..7556)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7624..7642
                     /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 ctaatattta agacttctta
     1801 atttcaaggc accccgggct ttactcgtaa agcttccggc gcgtatgttg tgtcgaccga
     1861 cctgcaggtg cagtaaggag gaaaaaaaaa tgaaatttgg aaactttttg cttacatacc
     1921 aacctcccca attttcccaa acagaggtaa tgaaacgttt ggttaaatta ggtcgcatct
     1981 ctgaggagtg tggttttgat accgtatggt tactggagca tcatttcacg gagtttggtt
     2041 tgcttggtaa cccttatgtc gctgctgcat atttacttgg cgcgactaaa aaattgaatg
     2101 taggaactgc cgctattgtt cttcccacag cccatccagt acgccaactt gaagatgtga
     2161 atttattgga tcaaatgtca aaaggacgat ttcggtttgg tatttgccga gggctttaca
     2221 acaaggactt tcgcgtattc ggcacagata tgaataacag tcgcgcctta gcggaatgct
     2281 ggtacgggct gataaagaat ggcatgacag agggatatat ggaagctgat aatgaacata
     2341 tcaagttcca taaggtaaaa gtaaaccccg cggcgtatag cagaggtggc gcaccggttt
     2401 atgtggtggc tgaatcagct tcgacgactg agtgggctgc tcaatttggc ctaccgatga
     2461 tattaagttg gattataaat actaacgaaa agaaagcaca acttgagctt tataatgaag
     2521 tggctcaaga atatgggcac gatattcata atatcgacca ttgcttatca tatataacat
     2581 ctgtagatca tgactcaatt aaagcgaaag agatttgccg gaaatttctg gggcattggt
     2641 atgattctta tgtgaatgct acgactattt ttgatgattc agaccaaaca agaggttatg
     2701 atttcaataa agggcagtgg cgtgactttg tattaaaagg acataaagat actaatcgcc
     2761 gtattgatta cagttacgaa atcaatcccg tgggaacgcc gcaggaatgt attgacataa
     2821 ttcaaaaaga cattgatgct acaggaatat caaatatttg ttgtggattt gaagctaatg
     2881 gaacagtaga cgaaattatt gcttccatga agctcttcca gtctgatgtc atgccatttc
     2941 ttaaagaaaa acaacgttcg ctattatatt atggcggtgg cggtagcggc ggtggcggta
     3001 gcggcggtgg cggtagcggc ggtggcggta gcaaatttgg attgttcttc cttaacttca
     3061 tcaattcaac aactgttcaa gaacagagta tagttcgcat gcaggaaata acggagtatg
     3121 ttgataagtt gaattttgaa cagattttag tgtatgaaaa tcatttttca gataatggtg
     3181 ttgtcggcgc tcctctgact gtttctggtt ttctgctcgg tttaacagag aaaattaaaa
     3241 ttggttcatt aaatcacatc attacaactc atcatcctgt ccgcatagcg gaggaagctt
     3301 gcttattgga tcagttaagt gaagggagat ttattttagg gtttagtgat tgcgaaaaaa
     3361 aagatgaaat gcattttttt aatcgcccgg ttgaatatca acagcaacta tttgaagagt
     3421 gttatgaaat cattaacgat gctttaacaa caggctattg taatccagat aacgattttt
     3481 atagcttccc taaaatatct gtaaatcccc atgcttatac gccaggcgga cctcggaaat
     3541 atgtaacagc aaccagtcat catattgttg agtgggcggc caaaaaaggt attcctctca
     3601 tctttaagtg ggatgattct aatgatgtta gatatgaata tgctgaaaga tataaagccg
     3661 ttgcggataa atatgacgtt gacctatcag agatagacca tcagttaatg atattagtta
     3721 actataacga agatagtaat aaagctaaac aagagacgcg tgcatttatt agtgattatg
     3781 ttcttgaaat gcaccctaat gaaaatttcg aaaataaact tgaagaaata attgcagaaa
     3841 acgctgtcgg aaattatacg gagtgtataa ctgcggctaa gttggcaatt gaaaagtgtg
     3901 gtgcgaaaag tgtattgctg tcctttgaac caatgaatga tttgatgagc caaaaaaatg
     3961 taatcaatat tgttgatgat aatattaaga agtaccacac ggaatatacc taaacggagc
     4021 caatgtacgc aaaaaacccc gcttcagcgg ggttttttcg ccaaaaaaaa ccccgccctg
     4081 tcaggggcgg ggtttttttt tagggcccta ctgactgtta caagcaaaga attaccttgt
     4141 ctggggaaat ttcgtcttca gtgtcaggga tagtgggcag cagagcagaa cgcgtcaggc
     4201 cccaaaactt ctggggagac atgtctttct tagtagccgt gaacttccag ccgatgtgag
     4261 atgcgcaaat tttgcattgt gcaacagttg cggctgcccc agggaaccaa gagtgctcgg
     4321 tagaaggacg accgataaga ttaagattgc atgctttata taccgtcagc gtttcgtgga
     4381 cgtagccatg aggattaacg tacgcagcca tcggcccgca aagagacaga ctaaagatct
     4441 cattcttcgt tgtaatttcc gtttcctggc attgcttaca gcacagacta gtgcacttat
     4501 tcataatgtc aagttcacaa cgcaagcgtt ggatagccga accaattttc aaaagttgga
     4561 tacgtaagac atcatcaatg ggaaggcacg cagccacgcg gtacgagaag tcaattgggt
     4621 tggaaggaag ggagtcgtct tttaagtttt cgtcccactc gcgtaattgt tttttaatac
     4681 ggtccataag cgtctctgcg tcgtacaacg aatataacca acgcggccag cttgtaaggt
     4741 tggcacaatg aaatttacgc ttctggtact tttgccacca cttgtagcta cattggtcct
     4801 cacgggacac tggcttcgac ggaaagattt gacatttatt taaggactcc aattgcactg
     4861 ctgacatagt agagggcagg acgcactctg gtaagatttg gaccttggcc tgctggattc
     4921 catcgctttg ggtacgcaat tccagaactt tgaagcgttg acgtccgatg gcttttactt
     4981 ttacaatttc gatcccgaag tcctgttctt cacgatatgc gtaaatctcg gctgtggttc
     5041 caaattgtgc ttcgcgttcc tggacattgc tgtaggctaa aactgcaaag gtgcggtcct
     5101 tctggatcaa attacgtacc atcgaaactt cttgcgggtg aaaaagttgc aaaggaagtg
     5161 tctggcccgg aatcaggatc atcatcactt gaggcagtac ggggataacc tggcaagagt
     5221 catcgtcgtg aagcgtacgg ccgtgaaact cctccatatc agcccccaga tatgtgtgac
     5281 ttgtaggcag gctggtatcg aaattgataa tgttaggctt ttttgcttct ttcgaatctt
     5341 gatcctcgac ttccatctca tcctcctcct cactctctgc tggcagcaac ggcagatggt
     5401 tccccatatt atgcgctgcg tcctgttgat ccccctctcc ggccttgtca tcgtcgtcct
     5461 tgtagtcgac atttgtctgt gacaaatcgc cttttaacag gtagtcgggc gaacactgta
     5521 aggctttcga aagggccaat aagttctccc cattaggctc tgtttcactg cgctcccact
     5581 gggaaatggc aacgttactc acacctacca tcttccccag agccgcttga cggatcttta
     5641 attttttgcg acgtgcgcga atgcgttctc ccatcagttg agtgtttgcg cttacccagg
     5701 agatcaatgg gtaacttcca cgtggttcgt ggcgcgagtg ataagcaaca ttcgtctggg
     5761 ataaatcgcc cttaagtaaa tagtcaggcg agcattgcag ggccttgcta agcgccagaa
     5821 ggttttcgcc gttcggctcc gtttcgctac gctcccattg agaaatggct acgttagata
     5881 caccgaccat tttccctaag gccgcctgac ggatcttaag cttcttacgg cgagcgcgga
     5941 tgcgctcacc cattaactgg gtgttcattt ttttttcctc cttactctag taaaagttaa
     6001 acaaaattat ttgtagaggg aaaccgttgt ggtctccctg aatatagata ccgagcctta
     6061 tgcatgcccg taaagttatc cagcaaccac tcatagacct agggcagcag atagggacga
     6121 cgtggtgtta gctgtgctct agaatcctca gcgagtttgt agaaacgcaa aaaggccatc
     6181 cgtcaggatg gccttctgct taatttgatg cctggcagtt tatggcgggc gtcctgcccg
     6241 ccaccctccg ggccgttgct tcgcaacgtt caaatccgct cccggcggat ttgtcctact
     6301 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc
     6361 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagac cccacactac
     6421 catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg accaccgcgc
     6481 tactgccgcc aggcaaattc tgttttatca gaccgcttct gcgttctgat ttaatctgta
     6541 tcaggctgaa aatcttctct catccgccaa aacagccatg cccatggcgt ttaccaatgc
     6601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgccca
     7201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7441 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7561 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     7621 aaaaataggc gtatcacgag gcccttaggc caaataggcc gt
//
