LOCUS       sequence-430206-        7639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid circuit displaying mmCRBN bait and driving expression of 
            luxAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7639)
  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 7639)
  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: pSD090A
FEATURES             Location/Qualifiers
     source          1..7639
                     /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             1888..2967
                     /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             3031..4011
                     /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"
     protein_bind    5364..5375
                     /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(5451..5474)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(6147..6174)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6266..6352)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6505..6522
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(join(6568..7359,7360..7428))
                     /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     7191..7210
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7429..7533)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7601..7619
                     /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 aaatttggaa actttttgct tacataccaa
     1921 cctccccaat tttcccaaac agaggtaatg aaacgtttgg ttaaattagg tcgcatctct
     1981 gaggagtgtg gttttgatac cgtatggtta ctggagcatc atttcacgga gtttggtttg
     2041 cttggtaacc cttatgtcgc tgctgcatat ttacttggcg cgactaaaaa attgaatgta
     2101 ggaactgccg ctattgttct tcccacagcc catccagtac gccaacttga agatgtgaat
     2161 ttattggatc aaatgtcaaa aggacgattt cggtttggta tttgccgagg gctttacaac
     2221 aaggactttc gcgtattcgg cacagatatg aataacagtc gcgccttagc ggaatgctgg
     2281 tacgggctga taaagaatgg catgacagag ggatatatgg aagctgataa tgaacatatc
     2341 aagttccata aggtaaaagt aaaccccgcg gcgtatagca gaggtggcgc accggtttat
     2401 gtggtggctg aatcagcttc gacgactgag tgggctgctc aatttggcct accgatgata
     2461 ttaagttgga ttataaatac taacgaaaag aaagcacaac ttgagcttta taatgaagtg
     2521 gctcaagaat atgggcacga tattcataat atcgaccatt gcttatcata tataacatct
     2581 gtagatcatg actcaattaa agcgaaagag atttgccgga aatttctggg gcattggtat
     2641 gattcttatg tgaatgctac gactattttt gatgattcag accaaacaag aggttatgat
     2701 ttcaataaag ggcagtggcg tgactttgta ttaaaaggac ataaagatac taatcgccgt
     2761 attgattaca gttacgaaat caatcccgtg ggaacgccgc aggaatgtat tgacataatt
     2821 caaaaagaca ttgatgctac aggaatatca aatatttgtt gtggatttga agctaatgga
     2881 acagtagacg aaattattgc ttccatgaag ctcttccagt ctgatgtcat gccatttctt
     2941 aaagaaaaac aacgttcgct attatattat ggcggtggcg gtagcggcgg tggcggtagc
     3001 ggcggtggcg gtagcggcgg tggcggtagc aaatttggat tgttcttcct taacttcatc
     3061 aattcaacaa ctgttcaaga acagagtata gttcgcatgc aggaaataac ggagtatgtt
     3121 gataagttga attttgaaca gattttagtg tatgaaaatc atttttcaga taatggtgtt
     3181 gtcggcgctc ctctgactgt ttctggtttt ctgctcggtt taacagagaa aattaaaatt
     3241 ggttcattaa atcacatcat tacaactcat catcctgtcc gcatagcgga ggaagcttgc
     3301 ttattggatc agttaagtga agggagattt attttagggt ttagtgattg cgaaaaaaaa
     3361 gatgaaatgc atttttttaa tcgcccggtt gaatatcaac agcaactatt tgaagagtgt
     3421 tatgaaatca ttaacgatgc tttaacaaca ggctattgta atccagataa cgatttttat
     3481 agcttcccta aaatatctgt aaatccccat gcttatacgc caggcggacc tcggaaatat
     3541 gtaacagcaa ccagtcatca tattgttgag tgggcggcca aaaaaggtat tcctctcatc
     3601 tttaagtggg atgattctaa tgatgttaga tatgaatatg ctgaaagata taaagccgtt
     3661 gcggataaat atgacgttga cctatcagag atagaccatc agttaatgat attagttaac
     3721 tataacgaag atagtaataa agctaaacaa gagacgcgtg catttattag tgattatgtt
     3781 cttgaaatgc accctaatga aaatttcgaa aataaacttg aagaaataat tgcagaaaac
     3841 gctgtcggaa attatacgga gtgtataact gcggctaagt tggcaattga aaagtgtggt
     3901 gcgaaaagtg tattgctgtc ctttgaacca atgaatgatt tgatgagcca aaaaaatgta
     3961 atcaatattg ttgatgataa tattaagaag taccacacgg aatataccta aacggagcca
     4021 atgtacgcaa aaaaccccgc ttcagcgggg ttttttcgcc aaaaaaaacc ccgccctgtc
     4081 aggggcgggg ttttttttta gggccctact gactgttaaa gacacaggat caccttatcg
     4141 ggactaattt catcctctgt ctctggaatt gtaggtaaaa gcgctgaacg ggttaacccc
     4201 cagaacttct ggggactcat atcctttttt gtggcggtaa atttccaacc aatgtgtgaa
     4261 gcacagattt tgcactgcgc aatggtccat gcgtaaccgg gaaaccaact gtggactgtg
     4321 ctgggacgac caataagatt caagttcgag gccttgtata cagtcagtgt ttcatggacg
     4381 tagccatgcg gattcacata agcagccata ggcccacaca atgacaagct gaagatttca
     4441 ttcttcgtgg taatttcagt ctcttggcat tgcttacagc acaatgacgt acatttattc
     4501 atgatatcca attcacagcg taaacgctga attgccgagc cgatttttaa taattgaata
     4561 cgtaaaacat catcgattgg aagacaagca gcaacgcggt acgagaagtc gataggattc
     4621 tcaggcaggg aatcgtcctt taagttttca tcccactcac gcaactgttt tttgatacga
     4681 tccatcaaag tttcagcgtc gtataacgaa tacaaccagc ggggccaaga agttaaattt
     4741 gcacagtgga atttgcgttt ttggtacttc tgccaccact tgcatgagta ttggtcctcc
     4801 cacgaaatgg gcttagacgg gaagacctgg catttgttca gagactccag ttgaactgca
     4861 ctcatggtgg aaggtaaaac acattctgga aggatttgta ccttggcttg ttgaatcccg
     4921 tcactctgtg tacgtaactc cagaaccttg aaacgttggc gtccgattgc tttgactttg
     4981 actacctcga taccgaactc ctgttcttcg cggtaggcat aaatttctgc tgttgtccca
     5041 aactgtgctt cgcgttcttg gacattgcta taagccaaaa ccgcgaaagt acggtctttc
     5101 tgaattaaat tacgaaccat cgatacttct tgggggtgcg ataattgaag aggaagggtc
     5161 tgccccggaa tcagaatcat aagaacttca ggtaatacgg gaattacttg gcatgaatca
     5221 tcgtcatgaa gcgtgcgccc gtgaaattct tccatatctg cacctaagta ggtatggcta
     5281 gtgggaagtg aggtgtcaaa attaatgatg ttcggtttgc gtgcttcttt ggagtcctga
     5341 tcctctactt ccatttcaat ttcgtcatcc tcatcctcgg aatctgccgg taacaacgga
     5401 agatgattgc ccatattgtg ggctgcatct tgttgatccc cctcccctgc cttgtcatcg
     5461 tcgtccttgt agtcaacatt cgaatcagag gtgccattga gcagccagtc aacacttacg
     5521 ccaagagctg acgcaagttc tggtaaaaag cgtggtcgct tagttttacc gttttcgagc
     5581 tgctctatag actgctgggt agtccccacc ttttgagcaa gttcagcctg gttaagtcca
     5641 agctgaattc ttttgctttt taccctgctg ctgatactag ctctaaccat gctaatcaat
     5701 ggatatttcc ctttgggctc aacgtgccca acaaatctaa cattcgaatc agaggtgcca
     5761 ttgagcagcc agtcaacact tacgccaaga gctgacgcaa gttctggtaa aaagcgtggt
     5821 cgcttagttt taccgttttc gagctgctct atagactgct gggtagtccc caccttttga
     5881 gcaagttcag cctggttaag tccaagctga attcttttgc tttttaccct gctgctgata
     5941 ctcatttttt tttcctcctt actctagtaa aagttaaaca aaattatttg tagagggaaa
     6001 ccgttgtggt ctccctgaat atagataccg agccttatgc atgcccgtaa agttatccag
     6061 caaccactca tagacctagg gcagcagata gggacgacgt ggtgttagct gtgctctaga
     6121 atcctcagcg agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttaa
     6181 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttcg
     6241 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     6301 aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg
     6361 cagttcccta ctctcgcatg gggagacccc acactaccat cggcgctacg gcgtttcact
     6421 tctgagttcg gcatggggtc aggtgggacc accgcgctac tgccgccagg caaattctgt
     6481 tttatcagac cgcttctgcg ttctgattta atctgtatca ggctgaaaat cttctctcat
     6541 ccgccaaaac agccatgccc atggcgttta ccaatgctta atcagtgagg cacctatctc
     6601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6661 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6721 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7081 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7141 agaatagtgt atgcggcgac cgagttgctc ttgcccagcg tcaatacggg ataataccgc
     7201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7441 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7501 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7561 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     7621 cttaggccaa ataggccgt
//
