LOCUS       sequence-175216-        5203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express Ubl4A tagged with VENUS at C terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5203)
  AUTHORS   Xu Y, Anderson DE, Ye Y
  TITLE     The HECT domain ubiquitin ligase HUWE1 targets unassembled soluble 
            proteins for degradation.
  JOURNAL   Cell Discov. 2016 Nov 8;2:16040. eCollection 2016.
  PUBMED    27867533
REFERENCE   2  (bases 1 to 5203)
  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: "Cell 
            Discov."; date: "2016-11-8"; volume: "2"; pages: "16040. eCollection
            2016"
COMMENT     Sequence Label: pVENUS-N1-Ubl4A
FEATURES             Location/Qualifiers
     source          1..5203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      79..667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     568..587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        842..1145
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1146..1349
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1300..1320
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1924..1933
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1930..1932,1933..1935,1936..2649)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELYK"
     primer_bind     complement(1975..1996)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2236..2255)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2772..2893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2809..2828)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2863..2882
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2900..3355)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3037..3058)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3249..3268
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3382..3486
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3488..3845
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3488..3679
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3511..3531)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3696..3831
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3880..4674
                     /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             3880..4674
                     /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(3934..3953)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4544..4563
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4862..4881)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4906..4953
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
       61 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      121 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      181 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      241 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      301 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      361 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      421 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      481 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      541 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      601 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      661 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      721 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
      781 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      841 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      901 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      961 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     1021 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     1081 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     1141 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1201 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1261 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1321 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
     1381 accggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcggccatg
     1441 cagctgacgg tgaaggcgct gcagggccgc gagtgcagcc tgcaggtgcc agaggacgag
     1501 ctggtgtcca cgctgaagca gctggtctcc gagaagctga acgtcccagt gcgccagcag
     1561 cggctgctgt tcaagggcaa ggccctggca gatgggaaac gactctcgga ttatagcatc
     1621 gggcccaact ccaagctcaa cctagtggtc aaacccctgg agaaggtgct actagaagaa
     1681 ggcgaggccc agaggctggc cgactcccca cccccgcagg tctggcagct gatctccaaa
     1741 gtcttggccc gccacttcag tgcggcagat gccagcaggg tcctggaaca gctacagagg
     1801 gattacgaga ggtccctgag tcgcctgacg ctggacgaca tcgaacggtt ggccagccgc
     1861 ttcctgcacc ctgaagtgac tgagacaatg gagaagggct tctccaaacg ggatccaccg
     1921 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1981 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2041 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     2101 tggcccaccc tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac
     2161 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2221 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2281 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2341 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     2401 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     2461 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2521 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2581 cacatggtcc tgctggaacg cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2641 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     2701 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2761 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2821 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2881 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2941 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3001 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3061 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3121 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3181 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3241 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3301 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     3361 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3421 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3481 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3661 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3781 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3841 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3901 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3961 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4021 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     4081 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4141 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4201 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4261 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4321 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4381 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     4441 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4501 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4561 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4621 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4681 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4741 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4801 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     4861 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4921 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4981 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     5041 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     5101 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     5161 atttttaatt taaaaggatc taggtgaaga tcctttttga taa
//
