LOCUS       sequence-442981-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell expression vector for IKZF1 mutant for CRBN cryoEM 
            pre-blotting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  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 7066)
  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: pSDC803_IKZF1_mutant
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(887..907)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(920..943)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(958..1049)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(1016..1033)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        1104..1213
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             1222..1899
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /label=DsRed-Monomer
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
                     VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
                     ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
                     GSQ"
     primer_bind     1620..1639
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1933..1952)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1977..2025
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    2215..2248
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_recomb     2252..3079
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             2312..2917
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     CDS             complement(2955..3079)
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     primer_bind     complement(3471..3488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3642..4230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3722..3741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4401..5192,5193..5261))
                     /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     5024..5043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5262..5366)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5434..5452
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5490..5512)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5612..5631
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5648..6103)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5785..5806)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5997..6016
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     misc_recomb     join(6250..7066,1..541)
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             join(6250..7066,1..525)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
ORIGIN
        1 cgtgtggcta tggcaaaatc gtcttcggaa gcaacttcta acgacgaggg ttgggacgac
       61 gacgataatc ggcctaataa agctaacacg cccgatgtta aatatgtcca agctttattt
      121 aacgtgttta cgtcgagtca attgtacact aacgacagtg atgaaagaaa tacaaaagcg
      181 cataatattt tgaacgacgt cgaaccttta ttacaaaaca aaacacaaac gaatatcgac
      241 aaagctagat tgctgctaca agatttggca agttttgtgg cgttgagcga aaatccatta
      301 gatagtccag ccatcggttc ggaaaaacaa cccttgtttg aaactaatcg aaacctattt
      361 tacaaatcta ttgaggattt aatatttaaa ttcagatata aagacgctga aaatcatttg
      421 attttcgctc taacatacca ccctaaagat tataaattta atgaattatt aaaatacgta
      481 caacaattgt ctgtaaatca acaacgcaca gaatctagcg cttaataaat gtactaataa
      541 cccggatcaa ttaattaagc ggccgtccgg gggtaccgtt gcacaaatga catttaaatg
      601 gtttctctcc tgagtgtaat tttatatgtc tcaagaggtt tcccttttga gtaaatgacg
      661 cattgcattg gttgcaagca aaaggtctct ctcctgttta ttacgagtgt gttctcaagt
      721 gaccggtgag agcatcacgg cggcggcaag cataattaca gaggtggcac ttgaagggct
      781 tctcacccga atgcaacttg atgtgtcgga gcaaattacc cttctgcgtg aaggaagcgt
      841 tacactgatt acacgcgaag ggacgttcgc ccgtcatgcg gccgcctccc tggaagtaga
      901 ggttctcgtc gaccgcggac ttgtcgtcat cgtctttgta gtccatggta gatccgtatt
      961 tataggtttt tttattacaa aactgttacg aaaacagtaa aatacttatt tatttgcgag
     1021 atggttatca ttttaattat ctccatgata tcagcaactc tattaatatt ccggagtagg
     1081 tcgcgaatcg atactagtat acggaccttt aattcaaccc aacacaatat attatagtta
     1141 aataagaatt attatcaaat catttgtata ttaattaaaa tactatactg taaattacat
     1201 tttatttaca atcactctac catggacaac accgaggacg tcatcaagga gttcatgcag
     1261 ttcaaggtgc gcatggaggg ctccgtgaac ggccactact tcgagatcga gggcgagggc
     1321 gagggcaagc cctacgaggg cacccagacc gccaagctgc aggtgaccaa gggcggcccc
     1381 ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa ggcctacgtg
     1441 aagcaccccg ccgacatccc cgactacatg aagctgtcct tccccgaggg cttcacctgg
     1501 gagcgctcca tgaacttcga ggacggcggc gtggtggagg tgcagcagga ctcctccctg
     1561 caggacggca ccttcatcta caaggtgaag ttcaagggcg tgaacttccc cgccgacggc
     1621 cccgtaatgc agaagaagac tgccggctgg gagccctcca ccgagaagct gtacccccag
     1681 gacggcgtgc tgaagggcga gatctcccac gccctgaagc tgaaggacgg cggccactac
     1741 acctgcgact tcaagaccgt gtacaaggcc aagaagcccg tgcagctgcc cggcaaccac
     1801 tacgtggact ccaagctgga catcaccaac cacaacgagg actacaccgt ggtggagcag
     1861 tacgagcacg ccgaggcccg ccactccggc tcccagtgac tcgagtaata gcgggagatg
     1921 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     1981 aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc
     2041 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattgggac caatacgccc
     2101 gcgtttcttc cttttcccca ccccaacccc caagttcggg tgaaggccca gggctcgcag
     2161 ccaacgtcgg ggcggcaagc cctgccatag ccactacggg tacgtttcaa ccagataact
     2221 tcgtataatg tatgctatac gaagttatat cagcaactat atattgatag acatttccag
     2281 tttgtgatat tagtttgtgc gtctcattac aatggctgtt atttttaaca acaaacaact
     2341 gctcgcagac aatagtatag aaaagggagg tgaactgttt ttgtttaacg gttcgtacaa
     2401 cattttggaa agttatgtta atccggtgct gctaaaaaat ggtgtaattg aactagaaga
     2461 agctgcgtac tatgccggca acatattgta caaaaccgac gatcccaaat tcattgatta
     2521 tataaattta ataattaaag caacacactc cgaagaacta ccagaaaata gcactgttgt
     2581 aaattacaga aaaactatgc gcagcggtac tatacacccc attaaaaaag acatatatat
     2641 ttatgacaac aaaaaattta ctctatacga tagatacata tatggatacg ataataacta
     2701 tgttaatttt tatgaggaga aaaatgaaaa agagaaggaa tacgaagaag aagacgacaa
     2761 ggcgtctagt ttatgtgaaa ataaaattat attgtcgcaa attaactgtg aatcatttga
     2821 aaatgatttt aaatattacc tcagcgatta taactacgcg ttttcaatta tagataatac
     2881 tacaaatgtt cttgttgcgt ttggtttgta tcgttaataa aaaacaaatt tgacatttat
     2941 aattgtttta ttattcaata attacaaata ggattgagac ccttgcagtt gccagcaaac
     3001 ggacagagct tgtcgacagt tttcattttt tggcagtttg gtggtttata cacatcttgg
     3061 gatttgatta aaagatgcat gacttcaccc acacacttgg agtcgtggtt gtagaaatgc
     3121 gtcagagcct tgattaaaca tcggttttca cacttggcac acgaaaccac gtttgcaatg
     3181 taacagttta gtataaacct cttcctgtac ataccgccgc gcggtacatt gttttctttc
     3241 aaatcgttca atattttttt tatacacgga ggcatattaa tcttgttgtt cagcatgtcg
     3301 acgacatttt tctttccttc tacgcattct ttcatacaaa tgtttctctt gctgcgtttt
     3361 ttaaaattgg aatttttaat ttcattaata gtgggcggag ccgttctgca ttaatgaatc
     3421 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3481 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3541 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3601 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3661 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3721 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3781 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3841 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3901 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3961 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4021 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4081 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4141 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4681 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5401 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5461 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5521 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5581 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     5641 caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5701 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5761 cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5821 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5881 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5941 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     6001 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     6061 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac
     6121 gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgacgac
     6181 agcgaagtgt ctcgccacta caatgctgta gtaaaaatga aaaggcttgt tatcggcgtg
     6241 cgggatcgat ccgagcttga gacaatctct ttacaataca attgcttaca ttgagcggtt
     6301 gttaaatatc ggcacggtaa acgatagcga aattacaatg cttatagcag atttttacga
     6361 tttgtattcc aattataata ttgaattgcc accaccccaa gcgctgcctc gaagcagacg
     6421 accttccgtt gtgcagccag cggcgcctgc gccggtgccc acaatcgtgc gcgaacaaac
     6481 taaaccagaa caaattatac cggcggcacc gccgccacca ccttctcccg tgcctaacat
     6541 tccagcgcct ccaccaccac caccaccatc gatgtctgaa ttgccgcccg ctccaccaat
     6601 gccgacggaa cctcaacccg ctgcaccttt agacgacaga caacaattgt tggaagctat
     6661 tagaaacgaa aaaaatcgca ctcgtctcag accggtcaaa ccaaaaacgg cgcccgaaac
     6721 cagtacaata gttgaggtgc cgactgtgtt gcctaaagag acatttgagc ctaaaccgcc
     6781 gtctgcatca ccgccaccac ctccgcctcc gcctccgccg ccagccccgc ctgcgcctcc
     6841 accgatggta gatttatcat cagctccacc accgccgcca ttagtagatt tgccgtctga
     6901 aatgttacca ccgcctgcac catcgctttc taacgtgttg tctgaattaa aatcgggcac
     6961 agttagattg aaacccgccc aaaaacgccc gcaatcagaa ataattccaa aaagctcaac
     7021 tacaaatttg atcgcggacg tgttagccga cacaattaat aggcgt
//
