LOCUS       sequence-271394-        4541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression 8xHis-Ub .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4541)
  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 4541)
  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: pCMV-8xHis-Ub
FEATURES             Location/Qualifiers
     source          1..4541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(233..250)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(260..487)
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             complement(260..487)
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             complement(497..514)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(543..746)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(572..592)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        747..1050
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1225..1813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1305..1324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2142..2189
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2214..2233
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2421..3215)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(2421..3215)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2532..2551)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3142..3161
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3250..3607)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3264..3399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3416..3607
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3564..3584
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3609..3713)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3740..4195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3827..3846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4037..4058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4202..4323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4213..4232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4267..4286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(4537..4541,1..223))
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             complement(join(4537..4541,1..223))
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
ORIGIN
        1 ctctgagacg gaggaccagg tgcagggtgg actctttctg gatgttgtag tcagacaggg
       61 tgcgtccatc ttccagctgt ttcccagcaa agatcaacct ctgctggtca ggagggatgc
      121 cttccttgtc ttggatcttt gccttgacat tctcaatggt gtcactcggc tccacttcga
      181 gagtgatggt cttaccagtc agggtcttca cgaagatctg catgcctcta gagtgatggt
      241 gatggtgatg gctagtcatc ccacctctga gacggaggac caggtgcagg gtggactctt
      301 tctggatgtt gtagtcagac agggtgcgtc catcttccag ctgtttccca gcaaagatca
      361 acctctgctg gtcaggaggg atgccttcct tgtcttggat ctttgccttg acattctcaa
      421 tggtgtcact cggctccact tcgagagtga tggtcttacc agtcagggtc ttcacgaaga
      481 tctgcatgcc tctagagtga tggtgatggt gatggctagc ggatctgacg gttcactaaa
      541 ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
      601 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
      661 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
      721 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
      781 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
      841 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
      901 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
      961 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     1021 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     1081 gaccccgtaa ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga
     1141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1801 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1861 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1921 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     1981 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     2041 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     2101 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     2161 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     2221 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     2281 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     2341 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     2401 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     2461 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     2521 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     2581 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     2641 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     2701 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     2761 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     2821 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     2881 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     2941 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     3001 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg
     3061 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     3121 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     3181 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     3241 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     3301 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     3361 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     3421 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     3481 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     3541 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc
     3601 gcctcaggac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3661 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3721 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3781 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3841 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3901 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3961 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4021 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4081 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4141 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacgcg
     4201 ttaagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4261 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4321 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     4381 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca gttatctaga
     4441 tccggtggat cccgggcccg cggtaccgtc gactgcagaa ttcgaagctt attaggtcag
     4501 tcagaattcc tgcagcccgg gggatccact agtcatccca c
//
