LOCUS       sequence-494589-        8035 bp    DNA     circular SYN 04-DEC-2025
DEFINITION  Expresses human UBC protein with an N-terminal cMyc tag enabling UBC
            pull down along with a blasticidin selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8035)
  AUTHORS   Wing CE, Fung HYJ, Kwanten B, Cagatay T, Niesman AB, Jacquemyn M, 
            Gharghabi M, Permentier B, Shakya B, Nandi R, Ready JM, Kashyap T, 
            Shacham S, Landesman Y, Lapalombella R, Daelemans D, Chook YM
  TITLE     SINE compounds activate exportin 1 degradation through an allosteric
            mechanism
  JOURNAL   Nat Chem Biol (2025) 21, 2002–2013.
  PUBMED    39416201
REFERENCE   2  (bases 1 to 8035)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol (2025) 21, 2002–2013."
COMMENT     Sequence Label: pLCKO-CMV-cMyc-UBC-IRES-Blasti
FEATURES             Location/Qualifiers
     source          1..8035
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             692..721
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             722..946
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="QIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFA
                     GKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             722..946
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="QIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFA
                     GKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     misc_feature    980..1554
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     1375..1394
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1587..1985
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    2001..2589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2054..2074)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2472..2483)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2661..2894
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2972..3093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3009..3028)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3063..3082
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3133..3268
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3289..3307)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3317..3334)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3317..3333)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3326..3348)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3475..3930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3562..3581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3772..3793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3956..4060
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4061..4129,4130..4921)
                     /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(4279..4298)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5092..5680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5581..5600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5834..5851
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5968..5989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6004..6009,6010..6027,6028..6034)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6042..6058
                     /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)."
     primer_bind     6047..6069
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6066..6082
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6066..6082
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6101..6121
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6103..6121
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6149..6375
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6376..6556
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6603..6728
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7221..7454
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7639..7683
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7870..7987
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagaccat ggaacagaag ctgatctccg aggaggacct
      721 tcagatcttc gtgaagaccc tgactggtaa gaccatcact ctcgaagtgg agccgagtga
      781 caccattgag aatgtcaagg caaagatcca agacaaggaa ggcatccctc ctgaccagca
      841 gaggttgatc tttgctggga aacagctgga agatggacgc accctgtctg actacaacat
      901 ccagaaagag tccaccctgc acctggtact ccgtctcaga ggtgggtgag gtaccggatc
      961 cggaattccg cccccccccc ccccctccct cccccccccc ctaacgttac tggccgaagc
     1021 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     1081 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     1141 ctttcccctc tcgccaaggg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     1201 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     1261 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     1321 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     1381 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     1441 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     1501 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc
     1561 cacaacccac aaggagacga ccttccatgg ccaagccttt gtctcaagaa gaatccaccc
     1621 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     1681 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     1741 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     1801 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     1861 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     1921 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     1981 gctgaacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     2041 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2101 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2161 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg cggcgtggtg tgcactgtgt
     2221 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2281 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2341 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2401 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     2461 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2521 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2581 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     2641 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     2701 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     2761 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     2821 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     2881 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
     2941 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     3001 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3061 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     3121 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3181 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3241 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     3301 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3361 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3421 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     3481 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3541 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3601 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3661 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3721 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3781 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3841 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3901 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     3961 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4021 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4081 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4141 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4201 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4261 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4321 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4381 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4441 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4501 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4561 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4621 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4681 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4741 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4801 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4861 tatggatgaa cgaaatagac agatcgctga gataggcgcc tcactgatta agcattggta
     4921 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4981 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5041 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5101 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5161 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5221 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5281 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5341 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5401 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5461 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5521 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5581 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5641 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5701 tttacggttc ctggtctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5761 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5821 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5881 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5941 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6001 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6061 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     6121 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     6181 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     6241 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
     6301 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
     6361 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6421 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6481 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6541 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     6601 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     6661 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     6721 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     6781 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     6841 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     6901 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     6961 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     7021 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     7081 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     7141 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     7201 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     7261 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     7321 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     7381 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     7441 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     7501 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     7561 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     7621 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     7681 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     7741 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     7801 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccaga
     7861 gagggcctat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     7921 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     7981 aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc tcgag
//
