LOCUS       sequence-169507-        8702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of full length HA tagged USPL1, catalytic 
            inactive variant C236S.
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-C236S-pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..8702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             910..936
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             916..942
                     /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(4255..4273)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4256..4273)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4293..4310)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4299..4523
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4569..4997
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4656..4675)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4866..4887
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5006..5026)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5011..5340
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5191..5326
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5253..5272
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5407..6201
                     /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(5461..5480)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6071..6090
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6375..6496
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6412..6431)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6466..6485
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6545..6561)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6545..6561)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6558..6580)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6569..6585
                     /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(6593..6599,6600..6617,6618..6623))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6638..6659
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6776..6793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6947..7535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7027..7046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7706..8497,8498..8566))
                     /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     8329..8348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8567..8671)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 atggcttcat atccttacga tgttccagac tacgcttccc ttggatccat gatggattct
      961 ccgaagattg gaaatggttt gccagtgatt ggaccaggga ctgatatagg gatatcttca
     1021 ctccacatgg tggggtattt gggaaaaaat tttgattcag ctaaagttcc atcagatgag
     1081 tattgccctg cttgtagaga gaagggaaag ttaaaagcct taaagactta ccgaattagt
     1141 tttcaagaat ctatcttttt gtgtgaggat ctgcagtgca tctatccttt gggctctaaa
     1201 tcacttaata acctaatttc tcctgatttg gaagaatgtc acactccaca taagcctcag
     1261 aaaaggaaga gcttagaaag cagctataag gattcacttc ttttagcaaa ttccaaaaag
     1321 actagaaatt atattgctat tgacggtgga aaagttttga acagcaaaca taatggagaa
     1381 gtatatgacg aaacctcgtc aaacttacct gatagtagtg gtcaacagaa tccaattagg
     1441 acagctgatt ccttggagcg gaatgagatt ttggaagctg atactgttga catggctact
     1501 acaaaagatc ctgctacagt tgatgtctct ggaactggca gaccttcccc tcaaaatgaa
     1561 ggatgtacat ctaaactgga aatgccactg gagagcaaat gtacatcatt tccccaggct
     1621 ttatgtgtcc agtggaaaaa tgcttatgct ctcagctggt tagactgtat cctgtcagct
     1681 ttggtgcact cggaagagtt aaagaacacc gtgactggac tgtgctcgaa ggaggaatct
     1741 atattctggc ggttgcttac aaaatataat caagcaaata cacttctata taccagtcaa
     1801 ttgagtggtg ttaaagatgg agattgtaaa aaacttacct cagaaatatt tgcagagata
     1861 gagacctgtc tgaatgaagt tagagatgaa atttttatta gccttcagcc ccagcttaga
     1921 tgcacattag gtgatatgga aagccctgtg tttgcatttc ccctgctctt aaaactagaa
     1981 acccacattg aaaagctctt cctatattct ttttcttggg actttgaatg ttcgcagtgt
     2041 ggacaccaat atcaaaacag gcatatgaag agtctggtca cctttacaaa tgtcatccct
     2101 gagtggcacc cacttaatgc tgcccatttt ggtccatgta acaattgcaa cagtaaatca
     2161 caaataagaa aaatggtatt agaaaaagta tctcccatat tcatgttgca ctttgtagaa
     2221 ggcttaccac agaatgactt gcagcactat gcatttcatt ttgaaggctg tctttatcag
     2281 ataacttctg taattcagta tcgagcaaat aatcatttta taacatggat tttagatgct
     2341 gatggaagtt ggctggaatg tgatgactta aaaggcccat gttctgaaag gcacaagaaa
     2401 tttgaagttc ctgcttcaga gatacatatt gttatttggg aaagaaaaat atcccaagtg
     2461 acagataaag aagctgcctg ccttccactt aaaaagacta atgaccaaca cgctctcagt
     2521 aatgagaaac cagtatcttt aacatcgtgt tctgtgggtg atgctgcctc agctgaaaca
     2581 gcctcagtaa ctcaccctaa agatatatca gttgcccctc gtactctttc acaggacaca
     2641 gctgtaactc atggagatca tttactttca ggtccaaaag gtttggttga caatatttta
     2701 cctctgacac ttgaagaaac tatccagaaa acagcctcag tttcacagtt aaattctgaa
     2761 gctttcctgt tagaaaataa acctgtagca gaaaatacag gaattctcaa aaccaatact
     2821 ttgctatcac aagaatcact aatggcttct tcagtatcag ctccatgtaa tgaaaagctt
     2881 attcaagacc aatttgtgga cataagtttt ccatcccaag ttgtaaatac aaacatgcag
     2941 tcagtacagc tgaatacaga agatactgta aatactaaat ctgtgaataa tactgatgct
     3001 actggtctta tacagggagt gaagtcagta gaaattgaga aggacgctca gttaaaacaa
     3061 ttccttacac caaaaactga acaattaaaa ccagaacgtg tcacatctca ggtatctaat
     3121 ttgaagaaaa aagaaactac agcagattct caaaccacaa catctaagtc attacagaat
     3181 cagtctctga aagaaaatca gaagaagcca tttgtgggaa gttgggttaa aggcttaata
     3241 agcaggggtg cttcttttat gccactctgt gtttcagctc ataatagaaa cactataact
     3301 gatttacaac cttcagttaa aggggtaaat aattttggtg gctttaaaac taaaggtata
     3361 aaccagaagg ccagccacgt atccaagaaa gctcgtaaga gtgcaagtaa gcctcctccc
     3421 atcagtaagc caccagcagg ccctccatcg tctaatggca cagctgccca cccacatgct
     3481 catgctgctt cagaagtttt ggaaaagtct ggaagcacct catgtggagc tcaactcaac
     3541 cacagttctt atgggaatgg tatttcttca gcaaaccatg aagacttggt ggaaggtcag
     3601 attcataaac ttcgtctaaa acttcgtaaa aagctaaagg cagaaaagaa gaaattagct
     3661 gctcttatgt cttccccgca aagcagaaca gttcgaagtg aaaatctaga acaggtgccc
     3721 caggatgggt ctccaaatga ttgtgaatca atagaggact tgttaaatga gctaccatat
     3781 ccaattgata ttgccagtga gtctgcatgc accactgttc ctggtgtttc cctgtacagt
     3841 agtcaaactc atgaagaaat tttagcggaa ttattgtctc ctacacctgt ttcaacagag
     3901 ctgtcagaaa atggggaagg tgactttagg tatttgggaa tgggagatag tcatatccca
     3961 ccaccagtac caagtgaatt caatgatgtt tcccagaaca cacatctgag acaggaccat
     4021 aattattgta gccccaccaa gaaaaatcca tgtgaagttc agccagactc tctgacaaat
     4081 aatgcctgcg ttagaacatt aaacttggag agtccgatga agactgatat tttcgatgag
     4141 tttttttcct cctcagcatt aaatgcttta gcaaatgaca cattagacct acctcatttc
     4201 gatgaatatc tgtttgagaa ttattgactc gagcatgcat ctagagggcc ctattctata
     4261 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     4321 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4381 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4441 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4501 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     4561 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4621 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4681 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4741 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4801 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4861 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4921 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4981 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5041 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5101 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5161 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5221 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5281 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5341 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5401 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5461 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5521 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5581 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5641 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5701 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5761 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5821 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5881 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5941 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6001 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6061 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6121 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6181 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6241 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6301 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6361 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6421 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6481 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6541 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6601 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6661 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6721 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6781 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6841 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6901 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6961 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7021 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7081 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7141 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7201 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7261 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7321 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7381 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7441 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7501 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7561 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7621 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7681 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7741 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7801 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7861 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7921 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7981 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8041 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8101 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8161 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8221 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8281 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8341 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8401 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8461 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8521 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8581 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8641 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8701 tc
//
