LOCUS       sequence-174905-        8702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of full length HA tagged USPL1, catalytic 
            inactive variant C236A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8702)
  AUTHORS   Schulz S, Chachami G, Kozaczkiewicz L, Winter U, Stankovic-Valentin 
            N, Haas P, Hofmann K, Urlaub H, Ovaa H, Wittbrodt J, Meulmeester E, 
            Melchior F
  TITLE     Ubiquitin-specific protease-like 1 (USPL1) is a SUMO isopeptidase 
            with essential, non-catalytic functions.
  JOURNAL   EMBO Rep. 2012 Oct;13(10):930-8. doi: 10.1038/embor.2012.125. Epub 
            2012 Aug 10.
  PUBMED    22878415
REFERENCE   2  (bases 1 to 8702)
  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"; doi: 
            "10.1038/embor.2012.125"; journalName: "EMBO Rep"; date: "2012-10"; 
            volume: "13"; issue: "10"; pages: "930-8"
COMMENT     Sequence Label: HA-USPL1-C236A-pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8702
                     /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"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             676..702
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             682..708
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(4021..4039)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4022..4039)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4059..4076)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4065..4289
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4335..4763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4422..4441)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4632..4653
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4772..4792)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4777..5106
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4957..5092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5019..5038
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5173..5967
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5227..5246)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5837..5856
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6141..6262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6178..6197)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6232..6251
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6311..6327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6311..6327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6324..6346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6335..6351
                     /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(6359..6365,6366..6383,6384..6389))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6404..6425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6542..6559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6713..7301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6793..6812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7472..8263,8264..8332))
                     /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     8095..8114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8333..8437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8512..8531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 acccaagctt
      661 ggtaccatgg cttcatatcc ttacgatgtt ccagactacg cttcccttgg atccatgatg
      721 gattctccga agattggaaa tggtttgcca gtgattggac cagggactga tatagggata
      781 tcttcactcc acatggtggg gtatttggga aaaaattttg attcagctaa agttccatca
      841 gatgagtatt gccctgcttg tagagagaag ggaaagttaa aagccttaaa gacttaccga
      901 attagttttc aagaatctat ctttttgtgt gaggatctgc agtgcatcta tcctttgggc
      961 tctaaatcac ttaataacct aatttctcct gatttggaag aatgtcacac tccacataag
     1021 cctcagaaaa ggaagagctt agaaagcagc tataaggatt cacttctttt agcaaattcc
     1081 aaaaagacta gaaattatat tgctattgac ggtggaaaag ttttgaacag caaacataat
     1141 ggagaagtat atgacgaaac ctcgtcaaac ttacctgata gtagtggtca acagaatcca
     1201 attaggacag ctgattcctt ggagcggaat gagattttgg aagctgatac tgttgacatg
     1261 gctactacaa aagatcctgc tacagttgat gtctctggaa ctggcagacc ttcccctcaa
     1321 aatgaaggat gtacatctaa actggaaatg ccactggaga gcaaatgtac atcatttccc
     1381 caggctttat gtgtccagtg gaaaaatgct tatgctctcg cttggttaga ctgtatcctg
     1441 tcagctttgg tgcactcgga agagttaaag aacaccgtga ctggactgtg ctcgaaggag
     1501 gaatctatat tctggcggtt gcttacaaaa tataatcaag caaatacact tctatatacc
     1561 agtcaattga gtggtgttaa agatggagat tgtaaaaaac ttacctcaga aatatttgca
     1621 gagatagaga cctgtctgaa tgaagttaga gatgaaattt ttattagcct tcagccccag
     1681 cttagatgca cattaggtga tatggaaagc cctgtgtttg catttcccct gctcttaaaa
     1741 ctagaaaccc acattgaaaa gctcttccta tattcttttt cttgggactt tgaatgttcg
     1801 cagtgtggac accaatatca aaacaggcat atgaagagtc tggtcacctt tacaaatgtc
     1861 atccctgagt ggcacccact taatgctgcc cattttggtc catgtaacaa ttgcaacagt
     1921 aaatcacaaa taagaaaaat ggtattagaa aaagtatctc ccatattcat gttgcacttt
     1981 gtagaaggct taccacagaa tgacttgcag cactatgcat ttcattttga aggctgtctt
     2041 tatcagataa cttctgtaat tcagtatcga gcaaataatc attttataac atggatttta
     2101 gatgctgatg gaagttggct ggaatgtgat gacttaaaag gcccatgttc tgaaaggcac
     2161 aagaaatttg aagttcctgc ttcagagata catattgtta tttgggaaag aaaaatatcc
     2221 caagtgacag ataaagaagc tgcctgcctt ccacttaaaa agactaatga ccaacacgct
     2281 ctcagtaatg agaaaccagt atctttaaca tcgtgttctg tgggtgatgc tgcctcagct
     2341 gaaacagcct cagtaactca ccctaaagat atatcagttg cccctcgtac tctttcacag
     2401 gacacagctg taactcatgg agatcattta ctttcaggtc caaaaggttt ggttgacaat
     2461 attttacctc tgacacttga agaaactatc cagaaaacag cctcagtttc acagttaaat
     2521 tctgaagctt tcctgttaga aaataaacct gtagcagaaa atacaggaat tctcaaaacc
     2581 aatactttgc tatcacaaga atcactaatg gcttcttcag tatcagctcc atgtaatgaa
     2641 aagcttattc aagaccaatt tgtggacata agttttccat cccaagttgt aaatacaaac
     2701 atgcagtcag tacagctgaa tacagaagat actgtaaata ctaaatctgt gaataatact
     2761 gatgctactg gtcttataca gggagtgaag tcagtagaaa ttgagaagga cgctcagtta
     2821 aaacaattcc ttacaccaaa aactgaacaa ttaaaaccag aacgtgtcac atctcaggta
     2881 tctaatttga agaaaaaaga aactacagca gattctcaaa ccacaacatc taagtcatta
     2941 cagaatcagt ctctgaaaga aaatcagaag aagccatttg tgggaagttg ggttaaaggc
     3001 ttaataagca ggggtgcttc ttttatgcca ctctgtgttt cagctcataa tagaaacact
     3061 ataactgatt tacaaccttc agttaaaggg gtaaataatt ttggtggctt taaaactaaa
     3121 ggtataaacc agaaggccag ccacgtatcc aagaaagctc gtaagagtgc aagtaagcct
     3181 cctcccatca gtaagccacc agcaggccct ccatcgtcta atggcacagc tgcccaccca
     3241 catgctcatg ctgcttcaga agttttggaa aagtctggaa gcacctcatg tggagctcaa
     3301 ctcaaccaca gttcttatgg gaatggtatt tcttcagcaa accatgaaga cttggtggaa
     3361 ggtcagattc ataaacttcg tctaaaactt cgtaaaaagc taaaggcaga aaagaagaaa
     3421 ttagctgctc ttatgtcttc cccgcaaagc agaacagttc gaagtgaaaa tctagaacag
     3481 gtgccccagg atgggtctcc aaatgattgt gaatcaatag aggacttgtt aaatgagcta
     3541 ccatatccaa ttgatattgc cagtgagtct gcatgcacca ctgttcctgg tgtttccctg
     3601 tacagtagtc aaactcatga agaaatttta gcggaattat tgtctcctac acctgtttca
     3661 acagagctgt cagaaaatgg ggaaggtgac tttaggtatt tgggaatggg agatagtcat
     3721 atcccaccac cagtaccaag tgaattcaat gatgtttccc agaacacaca tctgagacag
     3781 gaccataatt attgtagccc caccaagaaa aatccatgtg aagttcagcc agactctctg
     3841 acaaataatg cctgcgttag aacattaaac ttggagagtc cgatgaagac tgatattttc
     3901 gatgagtttt tttcctcctc agcattaaat gctttagcaa atgacacatt agacctacct
     3961 catttcgatg aatatctgtt tgagaattat tgactcgagc atgcatctag agggccctat
     4021 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     4081 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4141 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4201 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4261 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4321 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4381 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4441 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4501 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4561 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4621 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4681 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4741 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4801 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4861 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4921 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4981 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5041 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5101 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     5161 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5221 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5281 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5341 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5401 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5461 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5521 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5581 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5641 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5701 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5761 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5821 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5881 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5941 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6001 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6061 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6121 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     6181 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6241 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     6301 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6361 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6421 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6481 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6541 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6601 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6661 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6721 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6781 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6841 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6901 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6961 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7021 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7081 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7141 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7201 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7261 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7321 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7381 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7441 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7501 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7561 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7621 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7681 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7741 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7801 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7861 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7921 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7981 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8041 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8101 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8161 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8221 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8281 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8341 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8461 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     8521 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     8581 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     8641 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     8701 tt
//
