LOCUS       sequence-195150-        8990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of IKZF1 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 8990)
  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 8990)
  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-IKZF1-V2
FEATURES             Location/Qualifiers
     source          1..8990
                     /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(3688..3712)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3759..5411
                     /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(3825..3843)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5414..5638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5771..5790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5978..6099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5989..6008)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6043..6062
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6232..6336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6337..6405,6406..7197)
                     /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(6555..6574)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7368..7956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7857..7876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(8056..8075)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8247..8626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8627..8830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8781..8801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8827..8851
                     /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 tgctgatgag ggtcaagaca
     2281 tgtcccaagt ttcagggaag gaaagccccc ctgtaagcga tactccagat gagggcgatg
     2341 agcccatgcc gatccccgag gacctctcca ccacctcggg aggacagcaa agctccaaga
     2401 gtgacagagt cgtggccagt aatgttaaag tagagactca gagtgatgaa gagaatgggc
     2461 gtgcctgtga aatgaatggg gaagaatgtg cggaggattt acgaatgctt gatgcctcgg
     2521 gagagaaaat gaatggctcc cacagggacc aaggcagctc ggctttgtcg ggagttggag
     2581 gcattcgact tcctaacgga aaactaaagt gtgatatctg tgggatcatt tgcatcgggc
     2641 ccaatgtgct catggttcac aaaagaagcc acactggaga acggcccttc cagtgcaatc
     2701 agtgcggggc ctcattcacc cagaagggca acctgctccg gcacatcaag ctgcattccg
     2761 gggagaagcc cttcaaatgc cacctctgca actacgcctg ccgccggagg gacgccctca
     2821 ctggccacct gaggacgcac tccgtcatta aagaagaaac taatcacagt gaaatggcag
     2881 aagacctgtg caagatagga tcagagagat ctctcgtgct ggacagacta gcaagtaacg
     2941 tcgccaaacg taagagctct atgcctcaga aatttcttgg ggacaagggc ctgtccgaca
     3001 cgccctacga cagcagcgcc agctacgaga aggagaacga aatgatgaag tcccacgtga
     3061 tggaccaagc catcaacaac gccatcaact acctgggggc cgagtccctg cgcccgctgg
     3121 tgcagacgcc cccgggcggt tccgaggtgg tcccggtcat cagcccgatg taccagctgc
     3181 acaagccgct cgcggagggc accccgcgct ccaaccactc ggcccaggac agcgccgtgg
     3241 agaacctgct gctgctctcc aaggccaagt tggtgccctc ggagcgcgag gcgtccccga
     3301 gcaacagctg ccaagactcc acggacaccg agagcaacaa cgaggagcag cgcagcggtc
     3361 tcatctacct gaccaaccac atcgccccgc acgcgcgcaa cgggctgtcg ctcaaggagg
     3421 agcaccgcgc ctacgacctg ctgcgcgccg cctccgagaa ctcgcaggac gcgctccgcg
     3481 tggtcagcac cagcggggag cagatgaagg tgtacaagtg cgaacactgc cgggtgctct
     3541 tcctggatca cgtcatgtac accatccaca tgggctgcca cggcttccgt gatccttttg
     3601 agtgcaacat gtgcggctac cacagccagg accggtacga gttctcgtcg cacataacgc
     3661 gaggggagca ccgcttccac atgagctacc caactttctt gtacaaagtg gtgggtggag
     3721 gcggttcagg tggcggagga agcggaggcg gtggaagtat ggaagacgcc aaaaacataa
     3781 agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata
     3841 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
     3901 aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga
     3961 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
     4021 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
     4081 tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg
     4141 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa
     4201 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     4261 tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata
     4321 gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg
     4381 tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg
     4441 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
     4501 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     4561 ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg
     4621 tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     4681 atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg
     4741 ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag
     4801 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     4861 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag
     4921 gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     4981 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     5041 aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc
     5101 aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag
     5161 gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     5221 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     5281 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     5341 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga
     5401 tcgccgtgta agactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     5461 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     5521 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     5581 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     5641 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     5701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5881 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
     5941 tcctattctc tagagcggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     6001 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     6061 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     6121 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     6181 aaatgtggta aaatcgataa ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc
     6241 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6301 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6361 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6421 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6481 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6541 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6601 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6661 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6721 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6781 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6841 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6901 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6961 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7021 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7081 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7141 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7201 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7261 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7321 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7381 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7441 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7501 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7561 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7621 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7681 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7741 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7801 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7861 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7921 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7981 cggttcctgg ccttttgctg gccttttgct cacatggctc gacagatctc ccgatcccct
     8041 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     8101 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     8161 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     8221 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     8281 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     8341 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     8401 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     8461 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     8521 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     8581 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     8641 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     8701 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     8761 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     8821 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     8881 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
     8941 ggacctgcag gaattgatcc gcggccctag cgctaccgga ctcagatcca
//
