LOCUS       sequence-267709-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of USP7 UBL domains 1-3, with an N-terminal Myc
            tag and mutations in UBL2 (corresponding to DE758AA_D764A of the 
            full-length protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  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 7154)
  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_DED/AAA 535-888 USP7
FEATURES             Location/Qualifiers
     source          1..7154
                     /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(2745..2762)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2751..2975
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3021..3449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3108..3127)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3318..3339
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3458..3478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3463..3792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3643..3778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3705..3724
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3859..4653
                     /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(3913..3932)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4523..4542
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4827..4948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4864..4883)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4918..4937
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4997..5013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4997..5013)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5010..5032)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5021..5037
                     /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(5045..5051,5052..5069,5070..5075))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5090..5111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5228..5245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5399..5987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5479..5498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6158..6949,6950..7018))
                     /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     6781..6800
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7019..7123)
                     /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 cgatattcct cagcagttgg tggagcgatt acaagaagag aaaaggatcg aggctcagaa
     1021 gcggaaggag cggcaggaag cccatctcta tatgcaagtg cagatagtcg cagaggacca
     1081 gttttgtggc caccaaggga atgacatgta cgatgaagaa aaagtgaaat acactgtgtt
     1141 caaagtattg aagaactcct cgcttgctga gtttgttcag agcctctctc agaccatggg
     1201 atttccacaa gatcaaattc gattgtggcc catgcaagca aggagtaatg gaacaaaacg
     1261 accagcaatg ttagataatg aagccgacgg caataaaaca atgattgagc tcagtgataa
     1321 tgaaaaccct tggacaatat tcctggaaac agttgatccc gagctggctg ctagtggagc
     1381 gaccttaccc aagtttgata aagatcatga tgtaatgtta tttttgaaga tgtatgatcc
     1441 caaaacgcgg agcttgaatt actgtgggca tatctacaca ccaatatcct gtaaaatacg
     1501 tgacttgctc ccagttatgt gtgacagagc aggatttatt caagatacta gccttatcct
     1561 ctatgaggaa gttaaaccga atttaacaga gagaattcag gactatgacg tgtctcttga
     1621 taaagccctt gccgctctaa tggatggcgc catcatagta tttcagaagg atgaccctga
     1681 aaatgataac agtgaattac ccaccgcaaa ggagtatttc cgagatctct accaccgcgt
     1741 tgatgtcatt ttctgtgata aaacaatccc taatgatcct ggatttgtgg ttacgttatc
     1801 aaatagaatg aattattttc aggttgcaaa gacagttgca cagaggctca acacagatcc
     1861 aatgttgctg cagtttttca agtctcaagg ttatagggat ggcccaggta atcctcttag
     1921 acataattat gaaggtactt taagagatct tctacagttc ttcaagccta gacaacctaa
     1981 gaaactttac tatcagcagc ttaagatgaa aatcacagac ttttgaggat cccgaagttt
     2041 taaatgtata tggttaaaca gccaatttag ggaagaggaa ataacactat atccagacaa
     2101 gcatgggtgt gtccgggacc tgttagaaga atgtaaaaag gccgtggagc ttggggagaa
     2161 agcatcaggg aaacttaggc tgctagaaat tgtaagctac aaaatcattg gtgttcatca
     2221 agaagatgaa ctattagaat gtttatctcc tgcaacgagc cggacgtttc gaatagagga
     2281 aatccctttg gaccaggtgg acatagacaa agagaatgag atgcttgtca cagtggcgca
     2341 tttccacaaa gaggtcttcg gaacgttcgg aatcccgttt ttgctgagga tacaccaggg
     2401 cgagcatttt cgagaagtga tgaagcgaat ccagagcctg ctggacatcc aggagaagga
     2461 gtttgagaag tttaaatttg caattgtaat gatgggccga caccagtaca taaatgaaga
     2521 cgagtatgaa gtaaatttga aagactttga gccacagccc ggtaatatgt ctcatcctcg
     2581 gccttggcta gggctcgacc acttcaacaa agccccaaag aggagtcgct acacttacct
     2641 tgaaaaggcc attaaaatcc ataactgagg atccgaattc tgcagatatc cagcacagtg
     2701 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2761 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2821 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2881 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2941 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3001 gggctttagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3061 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3121 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3181 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3241 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3301 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3361 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3421 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3541 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3601 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3661 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3721 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3781 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3841 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3901 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3961 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4021 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4081 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4141 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4201 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4261 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4321 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4381 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4441 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4501 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4561 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4621 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4681 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4741 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4801 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4861 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4921 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4981 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5041 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5101 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5161 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5221 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5281 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5341 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5401 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5461 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5521 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5581 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5641 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5701 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5761 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5821 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5881 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5941 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6001 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6061 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6121 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6181 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6241 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6301 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6361 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6421 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6481 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6541 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6601 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6661 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6721 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6781 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6841 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6901 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6961 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7021 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7081 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7141 tgccacctga cgtc
//
