LOCUS       sequence-214232-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5109..5230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5146..5165)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5200..5219
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5237..5692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5374..5395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5586..5605
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5719..5823
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5825..6182
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5825..6016
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5848..5868)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6033..6168
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6217..7011
                     /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             6217..7011
                     /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(6271..6290)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6881..6900
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7199..7218)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7243..7290
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7619..8207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8108..8127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagaa cagaaggatc tctccttaac gcctttcacc
     1381 attaagagga aagcgatgga ggagctgagc gctgatgaga ttcgacggag gcgccttgca
     1441 cgacttgctg gtggacagac ctctcagcca accaccccac tcacctctcc ccagagggag
     1501 aaccctccgg ggcctcccat agcggcatca gccccaggac cctctcagag tcttggtctc
     1561 aatgtccaca acatgacccc agctacctcc ccaataggtg catcaggagt agcccatcga
     1621 agccagagca gtgaaggagt cagttctctc agcagctcgc cctctaatag ccttgaaacg
     1681 caatctcagt ctctctcacg ttcccagagc atggatatcg atggtgtctc atgtgagaaa
     1741 agcatgtccc aggtggatgt ggattcagga attgaaaaca tggaggttga tgaaaatgat
     1801 cgaagagaaa agcggagcct cagtgataag gagccttcct cgggccctga agtgtctgaa
     1861 gagcaggcct tacagctggt ctgtaagatc ttccgtgtct cttggaagga ccgggacaga
     1921 gatgtcatct ttctttcttc tctttctgca cagtttaagc agaacccaaa agaagtattc
     1981 tccgatttta aggacttgat tggccagatt ttaatggaag tgctaatgat gtccactcag
     2041 accagagatg aaaacccatt tgccagtctg acagccacat cacagccaat tgctgcagca
     2101 gcacggtcac cagacagaaa tctcttgcta aacactggct ccaatccagg aacaagcccc
     2161 atgttctgca gcgtggcttc ctttggtgcc agctctttgt ctagtttggg agcctctggt
     2221 ggagcaagta attgggattc ctacagtgac catttcacca ttgaaacctg caaagagaca
     2281 gatatgctga actacctcat cgagtgtttc gaccgagttg gaatagagga aaaaaaagca
     2341 ccaaagatgt gcagccagcc agcagtcagc cagcttctga gcaacatccg ctcacagtgc
     2401 atatcccata ctgctttagt actacaaggc tccctaacac agcccaggtc cttgcagcag
     2461 ccgtccttcc tagtgccgta tatgctgtgt aggaatctcc catatggctt cattcaggaa
     2521 ctggtgagaa ccactcacca ggatgaagaa gtgttcaagc agatatttat ccccatttta
     2581 caaggcctgg ctcttgctgc caaagagtgc tccctcgaca gtgactactt taaatacccc
     2641 ctcatggcac taggtgagct ctgtgaaacc aagtttggga agacacaccc tgtgtgcaat
     2701 ttggttgctt ctttgcggtt gtggttgccg aaatccttaa gtcctggctg tgggcgggag
     2761 ctgcagagac tctcttactt aggggctttc tttagcttct cagtctttgc agaagatgat
     2821 gttaaagtgg ttgaaaaata cttctcaggg cctgccatta ccctggaaaa cactcgtgtg
     2881 gttagccaat cattgcagca ttacttagag ctcggaaggc aagagctttt taagattctg
     2941 catagtattt tgttaaatgg cgaaacccgt gaggctgctc tcagttacat ggcggctgtc
     3001 gtcaatgcca atatgaagaa agcacagatg cagacagatg atagattggt gtctacagat
     3061 ggatttatgc tgaatttcct ttgggtactg cagcagctaa gtacaaaaat caagttagaa
     3121 acagttgatc ccacgtatat ttttcaccca agatgtcgga ttactcttcc caatgatgag
     3181 acgcgtgtga atgcaacgat ggaagatgtg aatgactggc tgactgaact ctatggcgat
     3241 cagcctccat tttctgagcc gaaattccct acggagtgct tctttctcac cctgcatgct
     3301 caccacctct ctattctgcc tagttgccgt cgctatatcc gcagactccg ggctatccgg
     3361 gagctcaata gaactgtaga agatttgaaa aataatgaaa gccaatggaa agattcccca
     3421 ctggcaacta gacaccgcga aatgctgaag cgctgtaaaa ctcagcttaa gaaactggta
     3481 cggtgcaagg cctgtgctga tgctggccta cttgacgaga gcttcctgag aagatgtctg
     3541 aatttttatg gccttctcat tcagctgctg ctccgcatcc tggaccccgc atatcccgat
     3601 ataacactgc ctttaaattc agatgtcccc aaggtatttg cagcgttgcc tgagttttat
     3661 gtagaagatg ttgcagaatt tttatttttt attgtacaat actctcccca ggcgctttat
     3721 gagccctgta ctcaggatat tgtgatgttc cttgttgtga tgttgtgcaa ccagaactac
     3781 atccgaaacc catatttggt ggccaaactg gtagaagtca tgtttatgac caaccctgct
     3841 gttcagccac gaacccagaa gttttttgaa atgattgaga accatcctct ctccaccaag
     3901 ttgttggtac cttccctgat gaagttttat acagatgttg agcataccgg agccaccagt
     3961 gagttttatg acaagttcac aattcgctat catattagca ccatttttaa aagcctttgg
     4021 caaaacatag ctcaccatgg cacctttatg gaggagttca actccgggaa gcagtttgtt
     4081 cgctatataa acatgttgat aaacgacacg acgtttttgc tcgatgaaag tctggagtct
     4141 ctgaagcgaa tccatgaagt gcaggaagag atgaagaaca aagaacagtg ggaccagttg
     4201 ccccgggatc agcagcaggc tcgtcagtct cagcttgctc aggatgagcg tgtgtcccgc
     4261 tcttacctcg ccctggccac cgaaaccgtg gacatgttcc acatcctcac gaagcaggtc
     4321 cagaagccct tcctcagacc ggagcttgga ccccgattgg ctgcaatgct gaactttaat
     4381 cttcagcaac tttgtggccc caagtgccgt gacctgaaag ttgaaaaccc tgagaaatac
     4441 ggctttgaac caaagaagct gttggaccaa ctgacggata tttacttaca gctggactgt
     4501 gctcggttcg cgaaagccat tgctgacgac cagagatcct acagtaagga attgtttgaa
     4561 gaagttattt caaagatgcg gaaggcaggg atcaaatcca caatagcaat agaaaaattt
     4621 aagctgctcg ccgagaaagt ggaggagata gtggccaaga acgcacgcgc agaaatcgac
     4681 tacagcgacg ctcctgatga gttcagagac cctctgatgg acaccctcat gacagacccc
     4741 gtgcggctgc cctctggcac catcatggac cgctccatca tcctgcggca cctgctcaac
     4801 tcccccacgg accccttcaa ccggcagacg ctgacagaga gcatgctgga accagtgcca
     4861 gaactgaaag agcagattca ggcgtggatg agagagaaac agaacagcga tcactaaacc
     4921 gttccgccgc ccaccctctg ctagacacag ccaaggccaa cgaggcaagc agaagcagcg
     4981 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     5041 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     5101 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5161 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5221 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     5281 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5341 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5401 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5461 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     5521 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5581 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5641 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     5701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5821 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5881 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6001 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     6061 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6121 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     6181 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     6241 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6301 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6361 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     6421 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6481 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6541 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6601 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6661 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6721 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     6781 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6841 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6901 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6961 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     7021 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     7081 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     7141 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     7201 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     7261 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     7321 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     7381 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     7441 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     7501 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7741 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     7801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8281 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
