LOCUS       sequence-246542-        5853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses UBE2QL1 with N-terminal Strep-HA tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  AUTHORS   Koerver L, Papadopoulos C, Liu B, Kravic B, Rota G, Brecht L, 
            Veenendaal T, Polajnar M, Bluemke A, Ehrmann M, Klumperman J, 
            Jaattela M, Behrends C, Meyer H
  TITLE     The ubiquitin-conjugating enzyme UBE2QL1 coordinates lysophagy in 
            response to endolysosomal damage.
  JOURNAL   EMBO Rep. 2019 Aug 21:e48014. doi: 10.15252/embr.201948014.
  PUBMED    31432621
REFERENCE   2  (bases 1 to 5853)
  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: "EMBO Rep. 
            2019 Aug 21:e48014. doi: 10.15252/embr.201948014."
COMMENT     Sequence Label: pcDNA5FRT/TO-Strep/HA-UBE2QL1
FEATURES             Location/Qualifiers
     source          1..5853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..477
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        478..681
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     632..652
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    683..701
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    704..722
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             927..953
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             984..1007
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1044..1067
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     protein_bind    1080..1104
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1606..1630)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1668..1685)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1674..1898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2031..2050)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2182..2229
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2237..3259
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2237..3259
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    3389..3510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3426..3445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3480..3499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3559..3575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3559..3575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3572..3594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3583..3599
                     /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)."
     promoter        complement(join(3607..3613,3614..3631,3632..3637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3652..3673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3790..3807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3961..4549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4041..4060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4720..5511,5512..5580))
                     /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     5343..5362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5581..5685)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5760..5779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
       61 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
      121 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      181 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      241 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      301 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      361 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      421 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      481 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      541 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      601 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      661 gaggtctata taagcagagc tctccctatc agtgatagag atctccctat cagtgataga
      721 gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt
      781 tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctagc gtttaaactt
      841 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc agatatccag
      901 cacagtggcg gccgctcgag accatgtacc catacgatgt tcccgactac gccggtaccg
      961 agctcggatc caccatggct agctggagcc acccgcagtt cgagaaaggt ggaggttccg
     1021 gaggtggatc gggaggtgga tcgtggagcc acccgcagtt cgaaaaagcg gccgatatca
     1081 caagtttgta caaaaaagca ggcttctcga gcatgaagga gctgcaggac atcgcgcgcc
     1141 ttagcgaccg cttcatctcc gtggagctgg tggacgagag cctgttcgac tggaacgtga
     1201 agctgcacca ggtggacaag gactcggtgc tgtggcagga catgaaggag accaacaccg
     1261 agttcatcct gctcaacctc accttccccg acaacttccc cttctcgccg cccttcatgc
     1321 gggtgctcag cccgcgcctg gagaacggct acgtgctgga cggcggcgcc atctgcatgg
     1381 agctgctcac gccgcgcggc tggtccagcg cctacaccgt ggaggccgtc atgcgccagt
     1441 tcgcagccag cctggtcaag ggccagggac ggatctgtag aaaagctggc aaatcaaaaa
     1501 agtccttcag tcgcaaggaa gctgaagcta cctttaagag tttggtgaag acgcatgaaa
     1561 aatatggttg ggtcaccccg cccgtgtccg acggctgagg aattcaccca gctttcttgt
     1621 acaaagtggt gacgtaagct aggggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     1681 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1741 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1801 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1861 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     1921 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1981 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2041 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2101 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2161 gggtgatggt tcacgtacct agaagttcct attccgaagt tcctattctc tagaaagtat
     2221 aggaacttcc ttggccaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     2281 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     2341 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     2401 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     2461 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     2521 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     2581 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     2641 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     2701 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     2761 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     2821 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     2881 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     2941 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     3001 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     3061 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     3121 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     3181 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     3241 tccgagggca aaggaatagc acgtactacg agatttcgat tccaccgccg ccttctatga
     3301 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     3361 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     3421 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3481 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     3541 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3601 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3661 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3721 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4381 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5701 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     5761 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     5821 gtcgctgagt agtgcgcgag caaaatttaa gct
//
