LOCUS       sequence-212941-        3496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Ubiquitin (1-76; F4A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3496)
  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 3496)
  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; F4A)
FEATURES             Location/Qualifiers
     source          1..3496
                     /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)"
     CDS             404..631
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIAVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     primer_bind     complement(777..795)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      791..838
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(883..901)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        969..1073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1074..1142,1143..1934)
                     /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(1292..1311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2105..2693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2594..2613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2847..2864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2879..3018
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2944..2963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3104..3126
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3120..3311)
                     /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 tcgccgtgaa
      421 gaccctgact ggtaagacca tcactctcga agtggagccg agtgacacca ttgagaatgt
      481 caaggcaaag atccaagaca aggaaggcat ccctcctgac cagcagaggt tgatctttgc
      541 tgggaaacag ctggaagatg gacgcaccct gtctgactac aacatccaga aagagtccac
      601 cctgcacctg gtcctccgtc tcagaggtgg gtgataaaag cttggtaccg agctcggatc
      661 cactagtaac ggccgccagt gtgctggaat tctgcagata tccatcacac tggcggccgc
      721 tcgagcagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
      781 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      841 gaaaggagga actatatccg gataattctt gaagacgaaa gggcctcgtg atacgcctat
      901 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
      961 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     1621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     2941 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     3001 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3061 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3121 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     3181 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     3241 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     3301 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     3361 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     3421 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     3481 cagggtcaat gccagc
//
