LOCUS       sequence-210744-        6996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of IKZF2-V2-V5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6996)
  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 6996)
  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: pcDNA3-IKZF2-V2-V5
FEATURES             Location/Qualifiers
     source          1..6996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        130..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     758..777
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        758..776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    806..830
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2331..2355)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2381..2422
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2462..2481)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2506..2554
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2756..3184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2843..2862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3053..3074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3193..3213)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3198..3527
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3378..3513
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3440..3459
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3594..4388
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3648..3667)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4258..4277
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4564..4685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4601..4620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4655..4674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4734..4750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4734..4750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4747..4769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4758..4774
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4782..4788,4789..4806,4807..4812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4827..4848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4965..4982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5136..5724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5216..5235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5895..6686,6687..6755))
                     /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     6518..6537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6756..6860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6935..6954)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
       61 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      121 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      721 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      781 cccaagctgg ctagttaagc tatcaacaag tttgtacaaa aaagttggca tggaaacaga
      841 ggccattgat ggctatataa cgtgtgacaa tgagctttca cccgaaaggg agcactccaa
      901 tatggcaatt gacctcacct caagcacacc caatggacag catgcctcac caagtcacat
      961 gacaagcaca aattcagtaa agctagaaat gcagagtgat gaagagtgtg acaggaaacc
     1021 cctgagccgt gaagatgaga tcaggggcca tgatgagggt agcagcctag aagaacccct
     1081 aattgagagc agcgaggtgg ctgacaacag gaaagtccag gagcttcaag gcgagggagg
     1141 aatccggctt ccgaatggtg aacgcccctt ccactgtaac cagtgtggag cttcttttac
     1201 tcagaagggc aaccttctga gacacataaa gttacactct ggagagaagc cgttcaaatg
     1261 tcctttctgt agctacgcct gtagaagaag ggacgccctc acaggacacc tcaggaccca
     1321 ttctgtgggt aaacctcaca agtgcaacta ctgtggacga agctacaagc agcgcagttc
     1381 actggaggag cacaaggaac gctgccacaa ctatctccag aatgtcagca tggaggctgc
     1441 tgggcaggtc atgagtcacc atgtacctcc tatggaagat tgtaaggaac aagagcctat
     1501 tatggacaac aatatttctc tggtgccttt tgagagacct gctgtcatag agaagctcac
     1561 ggggaatatg ggaaaacgta aaagctccac tccacaaaag tttgtggggg aaaagctcat
     1621 gcgattcagc tacccagata ttcactttga tatgaactta acatatgaga aggaggctga
     1681 gctgatgcag tctcatatga tggaccaagc catcaacaat gcaatcacct accttggagc
     1741 tgaggccctt caccctctga tgcagcaccc gccaagcaca atcgctgaag tggccccagt
     1801 tataagctca gcttattctc aggtctatca tccaaatagg atagaaagac ccattagcag
     1861 ggaaactgct gatagtcatg aaaacaacat ggatggcccc atctctctca tcagaccaaa
     1921 gagtcgaccc caggaaagag aggcctctcc cagcaatagc tgcctggatt ccactgactc
     1981 agaaagcagc catgatgacc accagtccta ccaaggacac cctgccttaa atcccaagag
     2041 gaaacaaagc ccagcttaca tgaaggagga tgtcaaagct ttggatacta ccaaggctcc
     2101 taagggctct ctgaaggaca tctacaaggt cttcaatgga gaaggagaac agattagggc
     2161 cttcaagtgt gagcactgcc gagtcctttt cctagaccat gtcatgtaca ccattcacat
     2221 gggttgccat ggctaccggg acccactgga atgcaacatc tgtggctaca gaagccagga
     2281 ccgttatgag ttttcatcac acattgttcg aggggagcac acattccact acccaacttt
     2341 cttgtacaaa gtggttgatc tagagggccc gcggttcgaa ggtaagccta tccctaaccc
     2401 tctcctcggt ctcgattcta cgcgtaccgg ttagtaatga gtttaaacgg gggaggctaa
     2461 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     2521 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     2581 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     2641 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     2701 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     2761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2821 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2881 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2941 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3061 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3121 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3181 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3361 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3421 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3481 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3541 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3601 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3661 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3721 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3781 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3841 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3901 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3961 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4021 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4081 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4141 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     4201 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4261 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4321 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4381 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     4441 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4501 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4561 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4621 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4681 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4741 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4801 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4861 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4921 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4981 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5041 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5221 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5281 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5341 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5401 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5461 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5521 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5581 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5641 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5701 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5761 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5821 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5881 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5941 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6001 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6061 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6121 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6181 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6241 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6301 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6361 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6421 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6481 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6541 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6601 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6661 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6721 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6781 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6841 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     6901 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6961 taagccagta tctgctccct gcttgtgtgt tggagg
//
