LOCUS       sequence-488951-        5567 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  Entry vector for Gateway with USP8 (C223A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5567)
  AUTHORS   Nie L, Wang C, Liu X, Teng H, Li S, Huang M, Feng X, Pei G, Hang Q, 
            Zhao Z, Gan B, Ma L, Chen J
  TITLE     USP7 substrates identified by proteomics analysis reveal the 
            specificity of USP7.
  JOURNAL   Genes Dev. 2022 Sep 1;36(17-18):1016-1030. doi: 
            10.1101/gad.349848.122. Epub 2022 Oct 27.
  PUBMED    36302555
REFERENCE   2  (bases 1 to 5567)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genes 
            Dev."; date: "2022-09-1"; volume: "36(17-18):1016-1030. doi"; pages:
            " 10.1101/gad.349848.122. Epub 2022 Oct 27"
COMMENT     Sequence Label: pDonr 201_USP7_C223S
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(3391..3490)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3543..3629
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3721..3748
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3883..4471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3963..3982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4614..5423)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5337..5356
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5415..5434
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    join(5546..5567,1..78)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tgatagtgac ctgttcgttg caacaaattg ataagcaatg ctttcttata atgccaactt
       61 tgtacaagaa agctgggtgt cagttatgga ttttaatggc cttttcaagg taagtgtagc
      121 gactcctctt tggggctttg ttgaagtggt cgagccctag ccaaggccga ggatgagaca
      181 tattaccggg ctgtggctca aagtctttca aatttacttc atactcgtct tcatttatgt
      241 actggtgtcg gcccatcatt acaattgcaa atttaaactt ctcaaactcc ttctcctgga
      301 tgtccagcag gctctggatt cgcttcatca cttctcgaaa atgctcgccc tggtgtatcc
      361 tcagcaaaaa cgggattccg aacgttccga agacctcttt gtggaaatgc gccactgtga
      421 caagcatctc attctctttg tctatgtcca cctggtccaa agggatttcc tctattcgaa
      481 acgtccggct cgttgcagga gataaacatt ctaatagttc atcttcttga tgaacaccaa
      541 tgattttgta gcttacaatt tctagcagcc taagtttccc tgatgctttc tccccaagct
      601 ccacggcctt tttacattct tctaacaggt cccggacaca cccatgcttg tctggatata
      661 gtgttatttc ctcttcccta aattggctgt ttaaccatat acatttaaaa cttcgcctgt
      721 tctcaaagtc tgtgattttc atcttaagct gctgatagta aagtttctta ggttgtctag
      781 gcttgaagaa ctgtagaaga tctcttaaag taccttcata attatgtcta agaggattac
      841 ctgggccatc cctataacct tgagacttga aaaactgcag caacattgga tctgtgttga
      901 gcctctgtgc aactgtcttt gcaacctgaa aataattcat tctatttgat aacgtaacca
      961 caaatccagg atcattaggg attgttttat cacagaaaat gacatcaacg cggtggtaga
     1021 gatctcggaa atactccttt gcggtgggta attcactgtt atcattttca gggtcatcct
     1081 tctgaaatac tatgatgtca ccatccatta gttcatcaag ggctttatca agagacacgt
     1141 catagtcctg aattctctct gttaaattcg gtttaacttc ctcatagagg ataaggctag
     1201 tatcttgaat aaatcctgct ctgtcacaca taactgggag caagtcacgt attttacagg
     1261 atattggtgt gtagatatgc ccacagtaat tcaagctccg cgttttggga tcatacatct
     1321 tcaaaaataa cattacatca tgatctttat caaacttggg taaggtcgct ccactagcag
     1381 ccagctcggg atcaactgtt tccaggaata ttgtccaagg gttttcatta tcactgagct
     1441 caatcattgt tttattgccg tcggcttcat tatctaacat tgcaggtcgt tttgttccat
     1501 tactccttgc ttgcatgggc cacaatcgaa tttgatcttg tggaaatccc atggtctgag
     1561 agaggctctg aacaaactca gcaagcgagg agttcttcaa tactttaaac acagtgtatt
     1621 tcactttttc ttcatcgtac atgtcattcc cttggtggcc acaaaactgg tcctctgcga
     1681 ctatctgcac ttgcatatag agatgggctt cctgccgctc cttccgcttc tgagcctcga
     1741 tccttttctc ttcttgtaat cgctccacca actgctgagg aatatcatgg tcggtgaccg
     1801 cctgtaaaac ttcactcagt tttgattccc tgatgtagac taacatgtaa gcattagtgc
     1861 agtgtcgaac agacaggtcg tcatcgtgac ccccataatt gtgctcaatt gcttcctctt
     1921 tagtacacct tgacaccacg tcgtcatcaa atttacacca tttgccatcc cctttggggt
     1981 ttagataaac cacataatgt ccaccatgat tatctccact atgaaccagg actgcatgaa
     2041 gaatataatt tgcagggtcc ttaggatctg ttttttgcaa aaattcatca agtggtaact
     2101 gctctgggaa ttcaaaccta tcattgatct tgatattttg gtccgtctga gggtcataca
     2161 taaatctcat cagttgtaga tgtaacactg gtggcaatgt taggaatttc acacctttct
     2221 ctgcttcctg taagccatgt tccccagcgt cgtatttatt gtccccatcg agctgttcta
     2281 ctgccacata atccacaaat gattcaaata tatttttctt tcctttgata cttagctgga
     2341 tatcataata atcttctctt ctatcagacc gatagtctac ttctttacac tggatatagg
     2401 acaccatttt gccgcggaat aatttgggta tggtgccctc tacacaggtg cctttcatct
     2461 tattttccac attatcgagc aacactcgac aaagctcctg aacatcatgt tgcatgaagc
     2521 tatctaaagt ttcccaccca aatgactttg ttaacttttt tgttcctaca ggtttatcac
     2581 tatgctgtaa ttcatagaac actctttgta atgctaaagg gacgctttta gacgaatcat
     2641 ccccctcggt tggcatcatg tacacagcct ttcgtagctg attcgtgaaa aataacgtct
     2701 gtagcaggct gttcatgtaa ctagtcgctc cctgattctt taagccgacg tagcctgtgt
     2761 gcttctttga atcccacgca actccatggg gagcatccgc ctgtacaaag acttcaaagg
     2821 taactttgtc atcatctata aatcctttct caggatcggt cacttcactc caggccataa
     2881 aattggaaaa tccccaatca ttttctttat ggaagaacaa atgactaata cgacgactga
     2941 acgacttttc atcatctctg taatttatta tcttcagcac tgcttgtgca tggcaagacc
     3001 atgacgtgga atcagattca gcattgcact ggagaaagaa tcctacgctt ttttggtgtg
     3061 gtctgtctgg ataaaagcgt ggcatcacca taatcttcca tggcagattt cgcacaaaac
     3121 acggagggct aaggaccgac tcactcagtc tgctgaagcg ctccacagtg aactgaaagg
     3181 ttgcctcgga gcgccaactg gtgtcatcct ccatgtcctc ctccgcggtg ttgtgtccat
     3241 cactcagggc cacattccca ttgatcacag ggttctgagt aattcttggt gggtcatctg
     3301 tatctcccgc ttccatctcc atgtcctcgg gctcgctcaa ctgctgctcg cccgctttct
     3361 gctgctgctg ctgctgctgg tggttcatga agcctgcttt tttgtacaaa gttggcatta
     3421 taaaaaagca ttgctcatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     3481 tcattatttg gggcccgaga tccatgctag cgttaacgcg agagtaggga actgccaggc
     3541 atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc tgttgtttgt
     3601 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgtgaagc
     3661 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaactaagc
     3721 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttcct ggctagcggt
     3781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagct tgcgccgtcc
     4561 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
     4621 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     4681 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     4741 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     4801 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     4861 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     4921 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     4981 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     5041 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     5101 aatacctgga atgctgtttt tccggggatc gcagtggtga gtaaccatgc atcatcagga
     5161 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     5221 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     5281 ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg
     5341 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgac
     5401 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     5461 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     5521 acgggccaga gctgcagctg gatggcaaat aatgatttta ttttgac
//
