LOCUS       sequence-251367-        3996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3996)
  AUTHORS   Meyer C, Garzia A, Morozov P, Molina H, Tuschl T
  TITLE     The G3BP1-Family-USP10 Deubiquitinase Complex Rescues Ubiquitinated 
            40S Subunits of Ribosomes Stalled in Translation from Lysosomal 
            Degradation.
  JOURNAL   Mol Cell. 2020 Mar 19;77(6):1193-1205.e5. doi: 
            10.1016/j.molcel.2019.12.024. Epub 2020 Jan 24.
  PUBMED    31981475
REFERENCE   2  (bases 1 to 3996)
  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: 
            "10.1016/j.molcel.2019.12.024"; journalName: "Mol Cell"; date: 
            "2020-03-19- 19"; volume: "77"; issue: "6"; pages: "1193-1205.e5"
COMMENT     Sequence Label: pENTR4_G3BP2
FEATURES             Location/Qualifiers
     source          1..3996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(37..64)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(156..242)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     291..313
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     305..322
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     306..322
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    338..437
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1890..1989)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2006..2025)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2007..2025)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2030..2046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2030..2046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2148..2167)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2159..2968
                     /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     complement(2226..2245)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3115..3703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3604..3623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3857..3874
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
       61 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
      121 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
      181 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
      241 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
      301 acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact
      361 gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt
      421 gtacaaaaaa gcaggcttca tggttatgga gaagcccagt ccgctgcttg tagggcggga
      481 gtttgtgagg caatattata ctttgctgaa taaagctccg gaatatttac acaggtttta
      541 tggcaggaat tcttcctatg ttcatggtgg agtagatgct agtggaaagc cccaggaagc
      601 tgtttatggc caaaatgata tacaccacaa agtattatct ctgaacttca gtgaatgtca
      661 tactaaaatt cgtcatgtgg atgctcatgc aaccttgagt gatggagtag ttgtccaggt
      721 catgggtttg ctgtctaaca gtggacaacc agaaagaaag tttatgcaaa cctttgttct
      781 ggctcctgaa ggatctgttc caaataaatt ttatgttcac aatgatatgt ttcgttatga
      841 agatgaagtg tttggtgatt ctgagcctga acttgatgaa gaatcagaag atgaagtaga
      901 agaggaacaa gaagaaagac aaccatctcc tgaacctgtg caagaaaatg ctaacagtgg
      961 ttactatgaa gctcaccctg tgactaatgg catagaggag cctttggaag aatcctctca
     1021 tgaacctgaa cctgagccag aatctgaaac aaagactgaa gagctgaaac cacaagtgga
     1081 ggagaagaac ttagaagaac tagaggagaa atctactact cctcctccgg cagaacctgt
     1141 ttctctgcca caagaaccac caaaggcttt ctcctgggct tcagtgacca gtaaaaacct
     1201 gcctcctagt ggtactgttt cttcctctgg aattccaccc catgttaaag caccagtctc
     1261 acagccaaga gtcgaagcta aaccagaagt tcaatctcag ccacctcgtg tgcgtgaaca
     1321 acgacctaga gaacgacctg gttttcctcc tagaggacca agaccaggca gaggagatat
     1381 ggaacagaat gactctgaca accgtagaat aattcgctat ccagatagtc atcaactttt
     1441 tgttggtaac ttgccacatg atattgatga aaatgagcta aaggaattct tcatgagttt
     1501 tggaaacgtt gtggaacttc gcatcaatac caagggtgtt gggggaaagc ttccaaattt
     1561 tggttttgtg gtttttgatg actctgaacc agttcagaga atcttaattg caaaaccgat
     1621 tatgtttcga ggggaagtac gtttaaatgt ggaagagaaa aaaacaagag ctgcaagaga
     1681 gcgagaaacc agaggtggtg gtgatgatcg cagggatatt aggcgcaatg atcgaggtcc
     1741 cggtggtcca cgtggaattg tgggtggtgg aatgatgcgt gatcgtgatg gaagaggacc
     1801 tcctccaagg ggtggcatgg cacagaaact tggctctgga agaggaaccg ggcaaatgga
     1861 gggccgcttc acaggacagc gtcgctgaga cccagctttc ttgtacaaag ttggcattat
     1921 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     1981 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2041 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2101 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2161 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2221 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2281 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2341 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2401 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2461 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2521 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2581 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2641 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2701 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2761 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     2821 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     2881 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     2941 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3001 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3061 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3121 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3181 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3241 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3301 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3361 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3421 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3481 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3541 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3601 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3661 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3721 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3781 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3841 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3901 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     3961 ctggaaagcg ggcagtgagc gcaacgcaat taatac
//
