LOCUS       sequence-223480-        8321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human UBE4B with N-terminal GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8321)
  AUTHORS   Hulsmann J, Kravic B, Weith M, Gstaiger M, Aebersold RH, Collins BC,
            Meyer H
  TITLE     AP-SWATH reveals direct involvement of VCP/p97 in integrated stress 
            response signaling through facilitating CReP/PPP1R15B degradation.
  JOURNAL   Mol Cell Proteomics. 2018 Mar 29. pii: RA117.000471. doi: 
            10.1074/mcp.RA117.000471.
  PUBMED    29599191
REFERENCE   2  (bases 1 to 8321)
  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"; journalName: "Mol Cell 
            Proteomics. 2018 Mar 29. pii: RA117.000471. doi: 
            10.1074/mcp.RA117.000471."
COMMENT     Sequence Label: pEGFP-UBE4B
FEATURES             Location/Qualifiers
     source          1..8321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        144..447
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        448..651
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     602..622
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      690..699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(696..698,699..701,702..1412)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(696..698,699..701,702..1412)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(741..762)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1002..1021)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1349..1370
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5192..5313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5229..5248)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5283..5302
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5320..5775)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5457..5478)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5669..5688
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5802..5906
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5908..6265
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5908..6099
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5931..5951)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6116..6251
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6300..7094
                     /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             6300..7094
                     /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(6354..6373)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6964..6983
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7282..7301)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7326..7373
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7702..8290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8191..8210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
       61 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
      121 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      181 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      241 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      301 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      361 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      421 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      481 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      541 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      601 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      661 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt
      721 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      781 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      841 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      901 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      961 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1021 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1081 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     1141 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     1201 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1261 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     1321 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1381 ggatcactct cggcatggac gagctgtaca agtccggact cagatctcga gaacagaagg
     1441 atctctcctt aacgcctttc accattaaga ggaaagcgat ggaggagctg agcgctgatg
     1501 agattcgacg gaggcgcctt gcacgacttg ctggtggaca gacctctcag ccaaccaccc
     1561 cactcacctc tccccagagg gagaaccctc cggggcctcc catagcggca tcagccccag
     1621 gaccctctca gagtcttggt ctcaatgtcc acaacatgac cccagctacc tccccaatag
     1681 gtgcatcagg agtagcccat cgaagccaga gcagtgaagg agtcagttct ctcagcagct
     1741 cgccctctaa tagccttgaa acgcaatctc agtctctctc acgttcccag agcatggata
     1801 tcgatggtgt ctcatgtgag aaaagcatgt cccaggtgga tgtggattca ggaattgaaa
     1861 acatggaggt tgatgaaaat gatcgaagag aaaagcggag cctcagtgat aaggagcctt
     1921 cctcgggccc tgaagtgtct gaagagcagg ccttacagct ggtctgtaag atcttccgtg
     1981 tctcttggaa ggaccgggac agagatgtca tctttctttc ttctctttct gcacagttta
     2041 agcagaaccc aaaagaagta ttctccgatt ttaaggactt gattggccag attttaatgg
     2101 aagtgctaat gatgtccact cagaccagag atgaaaaccc atttgccagt ctgacagcca
     2161 catcacagcc aattgctgca gcagcacggt caccagacag aaatctcttg ctaaacactg
     2221 gctccaatcc aggaacaagc cccatgttct gcagcgtggc ttcctttggt gccagctctt
     2281 tgtctagttt gggagcctct ggtggagcaa gtaattggga ttcctacagt gaccatttca
     2341 ccattgaaac ctgcaaagag acagatatgc tgaactacct catcgagtgt ttcgaccgag
     2401 ttggaataga ggaaaaaaaa gcaccaaaga tgtgcagcca gccagcagtc agccagcttc
     2461 tgagcaacat ccgctcacag tgcatatccc atactgcttt agtactacaa ggctccctaa
     2521 cacagcccag gtccttgcag cagccgtcct tcctagtgcc gtatatgctg tgtaggaatc
     2581 tcccatatgg cttcattcag gaactggtga gaaccactca ccaggatgaa gaagtgttca
     2641 agcagatatt tatccccatt ttacaaggcc tggctcttgc tgccaaagag tgctccctcg
     2701 acagtgacta ctttaaatac cccctcatgg cactaggtga gctctgtgaa accaagtttg
     2761 ggaagacaca ccctgtgtgc aatttggttg cttctttgcg gttgtggttg ccgaaatcct
     2821 taagtcctgg ctgtgggcgg gagctgcaga gactctctta cttaggggct ttctttagct
     2881 tctcagtctt tgcagaagat gatgttaaag tggttgaaaa atacttctca gggcctgcca
     2941 ttaccctgga aaacactcgt gtggttagcc aatcattgca gcattactta gagctcggaa
     3001 ggcaagagct ttttaagatt ctgcatagta ttttgttaaa tggcgaaacc cgtgaggctg
     3061 ctctcagtta catggcggct gtcgtcaatg ccaatatgaa gaaagcacag atgcagacag
     3121 atgatagatt ggtgtctaca gatggattta tgctgaattt cctttgggta ctgcagcagc
     3181 taagtacaaa aatcaagtta gaaacagttg atcccacgta tatttttcac ccaagatgtc
     3241 ggattactct tcccaatgat gagacgcgtg tgaatgcaac gatggaagat gtgaatgact
     3301 ggctgactga actctatggc gatcagcctc cattttctga gccgaaattc cctacggagt
     3361 gcttctttct caccctgcat gctcaccacc tctctattct gcctagttgc cgtcgctata
     3421 tccgcagact ccgggctatc cgggagctca atagaactgt agaagatttg aaaaataatg
     3481 aaagccaatg gaaagattcc ccactggcaa ctagacaccg cgaaatgctg aagcgctgta
     3541 aaactcagct taagaaactg gtacggtgca aggcctgtgc tgatgctggc ctacttgacg
     3601 agagcttcct gagaagatgt ctgaattttt atggccttct cattcagctg ctgctccgca
     3661 tcctggaccc cgcatatccc gatataacac tgcctttaaa ttcagatgtc cccaaggtat
     3721 ttgcagcgtt gcctgagttt tatgtagaag atgttgcaga atttttattt tttattgtac
     3781 aatactctcc ccaggcgctt tatgagccct gtactcagga tattgtgatg ttccttgttg
     3841 tgatgttgtg caaccagaac tacatccgaa acccatattt ggtggccaaa ctggtagaag
     3901 tcatgtttat gaccaaccct gctgttcagc cacgaaccca gaagtttttt gaaatgattg
     3961 agaaccatcc tctctccacc aagttgttgg taccttccct gatgaagttt tatacagatg
     4021 ttgagcatac cggagccacc agtgagtttt atgacaagtt cacaattcgc tatcatatta
     4081 gcaccatttt taaaagcctt tggcaaaaca tagctcacca tggcaccttt atggaggagt
     4141 tcaactccgg gaagcagttt gttcgctata taaacatgtt gataaacgac acgacgtttt
     4201 tgctcgatga aagtctggag tctctgaagc gaatccatga agtgcaggaa gagatgaaga
     4261 acaaagaaca gtgggaccag ttgccccggg atcagcagca ggctcgtcag tctcagcttg
     4321 ctcaggatga gcgtgtgtcc cgctcttacc tcgccctggc caccgaaacc gtggacatgt
     4381 tccacatcct cacgaagcag gtccagaagc ccttcctcag accggagctt ggaccccgat
     4441 tggctgcaat gctgaacttt aatcttcagc aactttgtgg ccccaagtgc cgtgacctga
     4501 aagttgaaaa ccctgagaaa tacggctttg aaccaaagaa gctgttggac caactgacgg
     4561 atatttactt acagctggac tgtgctcggt tcgcgaaagc cattgctgac gaccagagat
     4621 cctacagtaa ggaattgttt gaagaagtta tttcaaagat gcggaaggca gggatcaaat
     4681 ccacaatagc aatagaaaaa tttaagctgc tcgccgagaa agtggaggag atagtggcca
     4741 agaacgcacg cgcagaaatc gactacagcg acgctcctga tgagttcaga gaccctctga
     4801 tggacaccct catgacagac cccgtgcggc tgccctctgg caccatcatg gaccgctcca
     4861 tcatcctgcg gcacctgctc aactccccca cggacccctt caaccggcag acgctgacag
     4921 agagcatgct ggaaccagtg ccagaactga aagagcagat tcaggcgtgg atgagagaga
     4981 aacagaacag cgatcactaa accgttccgc cgcccaccct ctgctagaca cagccaaggc
     5041 caacgaggca agcagaagca gcggatccac cggatctaga taactgatca taatcagcca
     5101 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     5161 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     5221 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5281 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     5341 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     5401 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     5461 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     5521 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5581 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5641 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5701 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5761 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5821 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5881 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     5941 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6001 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6061 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     6121 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6181 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6241 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     6301 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     6361 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6421 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6481 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6541 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6601 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6661 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6721 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6781 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     6841 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6901 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6961 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     7021 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     7081 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     7141 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     7201 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     7261 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     7321 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     7381 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     7441 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     7501 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     7561 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7621 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7681 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7741 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7801 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7861 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7921 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7981 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8041 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     8101 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8161 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8221 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8281 cctatggaaa aacgccagca acgcggcctt tttacggttc c
//
