LOCUS       sequence-218957-        3447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human Ubiquitin (1-76; T66V, L67N).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3447)
  AUTHORS   Gladkova C, Schubert AF, Wagstaff JL, Pruneda JN, Freund SM, 
            Komander D
  TITLE     An invisible ubiquitin conformation is required for efficient 
            phosphorylation by PINK1.
  JOURNAL   EMBO J. 2017 Dec 15;36(24):3555-3572. doi: 10.15252/embj.201797876. 
            Epub 2017 Nov 13.
  PUBMED    29133469
REFERENCE   2  (bases 1 to 3447)
  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: 
            "10.15252/embj.201797876"; journalName: "EMBO J"; date: "2017-12-15-
            15"; volume: "36"; issue: "24"; pages: "3555-3572"
COMMENT     Sequence Label: Ubiquitin (1-76; T66V, L67N)
FEATURES             Location/Qualifiers
     source          1..3447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     324..343
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        324..342
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             374..396
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(728..746)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      742..789
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(834..852)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        920..1024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1025..1093,1094..1885)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1243..1262)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2056..2644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2545..2564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2798..2815
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2830..2969
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2895..2914)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3055..3077
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3071..3262)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
       61 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
      121 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
      181 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
      241 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
      301 ccgagatctc gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc
      361 ctctagaaat aattttgttt aactttaaga aggagatata catatgcaga tcttcgtgaa
      421 gaccctgact ggtaagacca tcactctcga agtggagccg agtgacacca ttgagaatgt
      481 caaggcaaag atccaagaca aggaaggcat ccctcctgac cagcagaggt tgatctttgc
      541 tgggaaacag ctggaagatg gacgcaccct gtctgactac aacatccaga aagagtccgt
      601 caatcacctg gtcctccgtc tcagaggtgg gtgataagaa ttctgcagat atccatcaca
      661 ctggcggccg ctcgagcaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
      721 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      781 ggttttttgc tgaaaggagg aactatatcc ggataattct tgaagacgaa agggcctcgt
      841 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      901 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      961 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1021 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1081 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1141 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1201 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1261 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1321 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1381 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1441 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1501 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1561 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1621 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1681 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1741 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1801 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1861 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1921 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1981 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2041 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2101 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2161 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2221 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2281 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2341 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2401 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2461 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2521 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2581 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2641 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2701 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2761 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2821 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     2881 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     2941 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3001 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3061 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     3121 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3181 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3241 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     3301 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     3361 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     3421 gaaaaatcac tcagggtcaa tgccagc
//
