LOCUS       sequence-195077-        9086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of IKZF3 fused to firefly luciferase with co-expression 
            of Renilla luciferase, both under IRES control. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9086)
  AUTHORS   Lu G, Middleton RE, Sun H, Naniong M, Ott CJ, Mitsiades CS, Wong KK,
            Bradner JE, Kaelin WG Jr
  TITLE     The myeloma drug lenalidomide promotes the cereblon-dependent 
            destruction of Ikaros proteins.
  JOURNAL   Science. 2014 Jan 17;343(6168):305-9. doi: 10.1126/science.1244917. 
            Epub 2013 Nov 29.
  PUBMED    24292623
REFERENCE   2  (bases 1 to 9086)
  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"; doi: 
            "10.1126/science.1244917"; journalName: "Science"; date: 
            "2014-01-17- 17"; volume: "343"; issue: "6168"; pages: "305-9"
COMMENT     Sequence Label: pIRIGF-IKZF3
FEATURES             Location/Qualifiers
     source          1..9086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(16..590,591..593,594..602)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             603..1538
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    join(1626..2200,2201..2203,2204..2212)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     protein_bind    2232..2256
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3784..3808)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3855..5507
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3921..3939)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5510..5734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5867..5886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6074..6195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6085..6104)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6139..6158
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6328..6432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6433..6501,6502..7293)
                     /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     complement(6651..6670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7464..8052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7953..7972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(8152..8171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8343..8722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8723..8926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8877..8897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8923..8947
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ccggtatgta atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt
       61 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
      121 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
      181 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
      241 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
      301 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
      361 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
      421 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
      481 tctggggcct cggtacacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
      541 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
      601 ccatgacttc gaaagtttat gatccagaac aaaggaaacg gatgataact ggtccgcagt
      661 ggtgggccag atgtaaacaa atgaatgttc ttgattcatt tattaattat tatgattcag
      721 aaaaacatgc agaaaatgct gttatttttt tacatggtaa cgcggcctct tcttatttat
      781 ggcgacatgt tgtgccacat attgagccag tagcgcggtg tattatacca gaccttattg
      841 gtatgggcaa atcaggcaaa tctggtaatg gttcttatag gttacttgat cattacaaat
      901 atcttactgc atggtttgaa cttcttaatt taccaaagaa gatcattttt gtcggccatg
      961 attggggtgc ttgtttggca tttcattata gctatgagca tcaagataag atcaaagcaa
     1021 tagttcacgc tgaaagtgta gtagatgtga ttgaatcatg ggatgaatgg cctgatattg
     1081 aagaagatat tgcgttgatc aaatctgaag aaggagaaaa aatggttttg gagaataact
     1141 tcttcgtgga aaccatgttg ccatcaaaaa tcatgagaaa gttagaacca gaagaatttg
     1201 cagcatatct tgaaccattc aaagagaaag gtgaagttcg tcgtccaaca ttatcatggc
     1261 ctcgtgaaat cccgttagta aaaggtggta aacctgacgt tgtacaaatt gttaggaatt
     1321 ataatgctta tctacgtgca agtgatgatt taccaaaaat gtttattgaa tcggacccag
     1381 gattcttttc caatgctatt gttgaaggtg ccaagaagtt tcctaatact gaatttgtca
     1441 aagtaaaagg tcttcatttt tcgcaagaag atgcacctga tgaaatggga aaatatatca
     1501 aatcgttcgt tgagcgagtt ctcaaaaatg aacaatgact cgaggtttaa actatagagt
     1561 acgagccatg ataactgcag gtcgagcatg catctagggc ggccaattcc gcccctctcc
     1621 ccccccccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     1681 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     1741 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     1801 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     1861 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     1921 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     1981 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     2041 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     2101 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     2161 cacggggacg tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccacaagga
     2221 gacgaccttc cacaagtttg tacaaaaaag ttggcatgga agatatacaa acaaatgcgg
     2281 aactgaaaag cactcaggag cagtctgtgc ccgcagaaag tgcagcggtt ttgaatgact
     2341 acagtttaac caaatctcat gaaatggaaa atgtggacag tggagaaggc ccagccaatg
     2401 aagatgaaga cataggagat gattcaatga aagtgaaaga tgaatacagt gaaagagatg
     2461 agaatgtttt aaagtcagaa cccatgggaa atgcagaaga gcctgaaatc ccttacagct
     2521 attcaagaga atataatgaa tatgaaaaca ttaagttgga gagacatgtt gtctcattcg
     2581 atagtagcag gccaaccagt ggaaagatga actgcgatgt gtgtggatta tcctgcatca
     2641 gcttcaatgt cttaatggtt cataagcgaa gccatactgg tgaacgccca ttccagtgta
     2701 atcagtgtgg ggcatctttt actcagaaag gtaacctcct ccgccacatt aaactgcaca
     2761 caggggaaaa accttttaag tgtcacctct gcaactatgc atgccaaaga agagatgcgc
     2821 tcacggggca tcttaggaca cattctgtgg agaaacccta caaatgtgag ttttgtggaa
     2881 ggagttacaa gcagagaagt tcccttgagg agcacaagga gcgctgccgt acatttcttc
     2941 agagcactga cccaggggac actgcaagtg cggaggcaag acacatcaaa gcagagatgg
     3001 gaagtgaaag agctctcgta ctggacagat tagcaagcaa tgtggcaaaa cgaaaaagct
     3061 caatgcctca gaaattcatt ggtgagaagc gccactgctt tgatgtcaac tataattcaa
     3121 gttacatgta tgagaaagag agtgagctca tacagacccg catgatggac caagccatca
     3181 ataacgccat cagctatctt ggcgccgaag ccctgcgccc cttggtccag acaccgcctg
     3241 ctcccacctc ggagatggtt ccagttatca gcagcatgta tcccatagcc ctcacccggg
     3301 ctgagatgtc aaacggtgcc cctcaagagc tggaaaagaa aagcatccac cttccagaga
     3361 agagcgtgcc ttctgagaga ggcctctctc ccaacaatag tggccacgac tccacggaca
     3421 ctgacagcaa ccatgaagaa cgccagaatc acatctatca gcaaaatcac atggtcctgt
     3481 ctcgggcccg caatgggatg ccacttctga aggaggttcc ccgctcttac gaactcctca
     3541 agcccccgcc catctgccca agagactccg tcaaagtgat caacaaggaa ggggaggtga
     3601 tggatgtgta tcggtgtgac cactgccgcg tcctcttcct ggactatgtg atgttcacga
     3661 ttcacatggg ctgccacggc ttccgtgacc ctttcgagtg taacatgtgt ggatatcgaa
     3721 gccatgatcg gtatgagttc tcgtctcaca tagccagagg agaacacaga gccctgctga
     3781 agtgcccaac tttcttgtac aaagtggtgg gtggaggcgg ttcaggtggc ggaggaagcg
     3841 gaggcggtgg aagtatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
     3901 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
     3961 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
     4021 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
     4081 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
     4141 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
     4201 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
     4261 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
     4321 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
     4381 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
     4441 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
     4501 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
     4561 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
     4621 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
     4681 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
     4741 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
     4801 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
     4861 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg
     4921 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
     4981 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
     5041 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
     5101 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
     5161 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
     5221 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
     5281 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
     5341 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
     5401 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
     5461 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaagac tgtgccttct
     5521 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5581 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5641 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     5701 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     5761 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5821 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5881 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5941 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6001 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga gcggccgctt
     6061 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     6121 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     6181 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     6241 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     6301 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6361 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6421 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6481 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6541 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6601 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6661 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6721 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6781 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6841 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6901 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6961 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7021 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7081 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7141 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7201 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7261 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7321 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7381 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7441 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7501 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7561 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7621 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7681 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7741 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7801 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7861 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7921 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7981 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8041 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8101 tttgctcaca tggctcgaca gatctcccga tcccctatgg tgcactctca gtacaatctg
     8161 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     8221 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     8281 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     8341 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     8401 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     8461 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     8521 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     8581 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     8641 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     8701 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     8761 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     8821 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     8881 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     8941 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
     9001 atccagcctc cgcggccggg aacggtgcat tggaacggac ctgcaggaat tgatccgcgg
     9061 ccctagcgct accggactca gatcca
//
