LOCUS       sequence-267711-        8756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of N-terminally Myc-tagged USP7, with mutations
            in the TRAF-like domain (DW164AA) and UBL2 (DE758AA_D764A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8756)
  AUTHORS   Ashton NW, Valles GJ, Jaiswal N, Bezsonova I, Woodgate R
  TITLE     DNA Polymerase iota Interacts with Both the TRAF-like and UBL1-2 
            Domains of USP7.
  JOURNAL   J Mol Biol. 2021 Jan 22;433(2):166733. doi: 
            10.1016/j.jmb.2020.166733. Epub 2020 Dec 3.
  PUBMED    33279577
REFERENCE   2  (bases 1 to 8756)
  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: "J Mol 
            Biol."; date: "2021-01-22"; volume: "433(2):166733. doi"; pages: " 
            10.1016/j.jmb.2020.166733. Epub 2020 Dec 3"
COMMENT     Sequence Label: pcDNA3.1-N-Myc_DW164AA + DED/AAA USP7
FEATURES             Location/Qualifiers
     source          1..8756
                     /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"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             926..955
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4347..4364)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4353..4577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4623..5051
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4710..4729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4920..4941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5060..5080)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5065..5394
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5245..5380
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5307..5326
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5461..6255
                     /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(5515..5534)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6125..6144
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6429..6550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6466..6485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6520..6539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6599..6615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6599..6615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6612..6634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6623..6639
                     /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(6647..6653,6654..6671,6672..6677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6692..6713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6830..6847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7001..7589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7081..7100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7760..8551,8552..8620))
                     /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     8383..8402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8621..8725)
                     /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 gctggctagc
      901 gtttaaactt aagcttgcca ccatggagca gaaactcatc tctgaagagg atctgggtac
      961 catgaaccac cagcagcagc agcagcagca gaaagcgggc gagcagcagt tgagcgagcc
     1021 cgaggacatg gagatggaag cgggagatac agatgaccca ccaagaatta ctcagaaccc
     1081 tgtgatcaat gggaatgtgg ccctgagtga tggacacaac accgcggagg aggacatgga
     1141 ggatgacacc agttggcgct ccgaggcaac ctttcagttc actgtggagc gcttcagcag
     1201 actgagtgag tcggtcctta gccctccgtg ttttgtgcga aatctgccat ggaagattat
     1261 ggtgatgcca cgcttttatc cagacagacc acaccaaaaa agcgtaggat tctttctcca
     1321 gtgcaatgct gaatctgatt ccacgtcatg gtcttgccat gcacaagcag tgctgaagat
     1381 aataaattac agagatgatg aaaagtcgtt cagtcgtcgt attagtcatt tgttcttcca
     1441 taaagaaaat gccgccggat tttccaactt tatggcctgg agtgaagtga ccgatcctga
     1501 gaaaggattt atcgatgatg acaaagttac ctttgaagtc tttgtacagg cggatgctcc
     1561 ccatggagtt gcgtgggatt caaagaagca caccggttac gtcggcttaa agaatcaggg
     1621 agcgacttgt tacatgaaca gcctgctaca gacgttattt ttcacgaatc agctacgaaa
     1681 ggctgtgtac atgatgccaa ccgaggggga tgattcgtct aaaagcgtcc ctttagcatt
     1741 acaaagagtg ttctatgaat tacagcatag tgataaacct gtaggaacaa aaaagttaac
     1801 aaagtcattt gggtgggaaa ctttagatag cttcatgcaa catgatgttc aggagctttg
     1861 tcgagtgttg ctcgataatg tggaaaataa gatgaaaggc acctgtgtag agggcaccat
     1921 acccaaatta ttccgcggca aaatggtgtc ctatatccag tgtaaagaag tagactatcg
     1981 gtctgataga agagaagatt attatgatat ccagctaagt atcaaaggaa agaaaaatat
     2041 atttgaatca tttgtggatt atgtggcagt agaacagctc gatggggaca ataaatacga
     2101 cgctggggaa catggcttac aggaagcaga gaaaggtgtg aaattcctaa cattgccacc
     2161 agtgttacat ctacaactga tgagatttat gtatgaccct cagacggacc aaaatatcaa
     2221 gatcaatgat aggtttgaat tcccagagca gttaccactt gatgaatttt tgcaaaaaac
     2281 agatcctaag gaccctgcaa attatattct tcatgcagtc ctggttcata gtggagataa
     2341 tcatggtgga cattatgtgg tttatctaaa ccccaaaggg gatggcaaat ggtgtaaatt
     2401 tgatgacgac gtggtgtcaa ggtgtactaa agaggaagca attgagcaca attatggggg
     2461 tcacgatgac gacctgtctg ttcgacactg cactaatgct tacatgttag tctacatcag
     2521 ggaatcaaaa ctgagtgaag ttttacaggc ggtcaccgac catgatattc ctcagcagtt
     2581 ggtggagcga ttacaagaag agaaaaggat cgaggctcag aagcggaagg agcggcagga
     2641 agcccatctc tatatgcaag tgcagatagt cgcagaggac cagttttgtg gccaccaagg
     2701 gaatgacatg tacgatgaag aaaaagtgaa atacactgtg ttcaaagtat tgaagaactc
     2761 ctcgcttgct gagtttgttc agagcctctc tcagaccatg ggatttccac aagatcaaat
     2821 tcgattgtgg cccatgcaag caaggagtaa tggaacaaaa cgaccagcaa tgttagataa
     2881 tgaagccgac ggcaataaaa caatgattga gctcagtgat aatgaaaacc cttggacaat
     2941 attcctggaa acagttgatc ccgagctggc tgctagtgga gcgaccttac ccaagtttga
     3001 taaagatcat gatgtaatgt tatttttgaa gatgtatgat cccaaaacgc ggagcttgaa
     3061 ttactgtggg catatctaca caccaatatc ctgtaaaata cgtgacttgc tcccagttat
     3121 gtgtgacaga gcaggattta ttcaagatac tagccttatc ctctatgagg aagttaaacc
     3181 gaatttaaca gagagaattc aggactatga cgtgtctctt gataaagccc ttgccgctct
     3241 aatggatggc gccatcatag tatttcagaa ggatgaccct gaaaatgata acagtgaatt
     3301 acccaccgca aaggagtatt tccgagatct ctaccaccgc gttgatgtca ttttctgtga
     3361 taaaacaatc cctaatgatc ctggatttgt ggttacgtta tcaaatagaa tgaattattt
     3421 tcaggttgca aagacagttg cacagaggct caacacagat ccaatgttgc tgcagttttt
     3481 caagtctcaa ggttataggg atggcccagg taatcctctt agacataatt atgaaggtac
     3541 tttaagagat cttctacagt tcttcaagcc tagacaacct aagaaacttt actatcagca
     3601 gcttaagatg aaaatcacag actttgagaa caggcgaagt tttaaatgta tatggttaaa
     3661 cagccaattt agggaagagg aaataacact atatccagac aagcatgggt gtgtccggga
     3721 cctgttagaa gaatgtaaaa aggccgtgga gcttggggag aaagcatcag ggaaacttag
     3781 gctgctagaa attgtaagct acaaaatcat tggtgttcat caagaagatg aactattaga
     3841 atgtttatct cctgcaacga gccggacgtt tcgaatagag gaaatccctt tggaccaggt
     3901 ggacatagac aaagagaatg agatgcttgt cacagtggcg catttccaca aagaggtctt
     3961 cggaacgttc ggaatcccgt ttttgctgag gatacaccag ggcgagcatt ttcgagaagt
     4021 gatgaagcga atccagagcc tgctggacat ccaggagaag gagtttgaga agtttaaatt
     4081 tgcaattgta atgatgggcc gacaccagta cataaatgaa gacgagtatg aagtaaattt
     4141 gaaagacttt gagccacagc ccggtaatat gtctcatcct cggccttggc tagggctcga
     4201 ccacttcaac aaagccccaa agaggagtcg ctacacttac cttgaaaagg ccattaaaat
     4261 ccataactga ggatccgaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     4321 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4381 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4441 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4501 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4561 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4621 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4681 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4741 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4801 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4861 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4921 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4981 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5041 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5101 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5161 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5221 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5281 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5341 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5401 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5461 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5521 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5581 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5641 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5701 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5761 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5821 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5881 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5941 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6001 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6061 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6121 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6181 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6241 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6301 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6361 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6421 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6481 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6541 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     6601 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6841 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6901 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6961 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7021 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7081 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7141 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7201 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7261 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7321 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7381 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7441 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7501 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7561 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7621 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7681 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7741 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7861 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7921 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8281 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8641 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8701 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
