LOCUS       sequence-267705-        7130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a mutant USP7 TRAF-like domain (DW164AA), 
            with an N-terminal Myc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  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 7130)
  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 1-205 USP7
FEATURES             Location/Qualifiers
     source          1..7130
                     /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(2721..2738)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2727..2951
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2997..3425
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3084..3103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3294..3315
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3434..3454)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3439..3768
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3619..3754
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3681..3700
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3835..4629
                     /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(3889..3908)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4499..4518
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4803..4924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4840..4859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4894..4913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4973..4989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4973..4989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4986..5008)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4997..5013
                     /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(5021..5027,5028..5045,5046..5051))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5066..5087
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5204..5221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5375..5963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5455..5474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6134..6925,6926..6994))
                     /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     6757..6776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6995..7099)
                     /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 gcgtggtgag gatccaagca 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 atgaggatcc gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc
     2701 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2761 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2821 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2881 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2941 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3001 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3061 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3121 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3181 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3301 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3361 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3421 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3481 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3601 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3661 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3721 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3781 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3841 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3901 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3961 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4021 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4081 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4141 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4201 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4261 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4321 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4381 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4441 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4501 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4561 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4621 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4681 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4741 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4801 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4861 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4921 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4981 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5041 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5101 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5161 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5221 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5281 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5341 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5401 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5461 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5521 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5581 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5641 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5701 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5761 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5821 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5881 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5941 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6001 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6061 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6121 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6181 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6241 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6301 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6361 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6421 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6481 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6541 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6601 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6661 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6721 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6781 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6841 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6901 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6961 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7021 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7081 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
