LOCUS       sequence-452284-        4029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of yeast Ubc4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4029)
  AUTHORS   Bashore C, Dambacher CM, Goodall EA, Matyskiela ME, Lander GC, 
            Martin A
  TITLE     Ubp6 deubiquitinase controls conformational dynamics and substrate 
            degradation of the 26S proteasome.
  JOURNAL   Nat Struct Mol Biol. 2015 Sep;22(9):712-9. doi: 10.1038/nsmb.3075. 
            Epub 2015 Aug 24.
  PUBMED    26301997
REFERENCE   2  (bases 1 to 4029)
  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: "10.1038/nsmb.3075";
            journalName: "Nat Struct Mol Biol"; date: "2015-09"; volume: "22"; 
            issue: "9"; pages: "712-9"
COMMENT     Sequence Label: pAM236 Ubc4
FEATURES             Location/Qualifiers
     source          1..4029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     176..193
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    208..347
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(273..292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     433..455
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(449..640)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1149..1168
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1149..1167
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1199..1221
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1678..1695
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1748..1766)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1762..1809
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1846..2301
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1933..1952)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2143..2164
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2327..2431
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2432..2500,2501..3292)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2650..2669)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3463..4029,1..22)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3952..3971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
       61 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      121 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
      181 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
      241 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
      301 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
      361 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
      421 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
      481 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
      541 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
      601 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
      661 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
      721 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
      781 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
      841 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
      901 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
      961 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     1021 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     1081 acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag atctcgatcc
     1141 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gaaataattt
     1201 tgtttaactt taagaaggag atataccatg agttcttcta aacgtattgc taaagaacta
     1261 agtgatctag aaagggatcc acctacttca tgttcagccg gtccagtcgg cgatgatcta
     1321 tatcactggc aagcatccat catgggacct gccgattccc catacgccgg cggtgttttc
     1381 ttcttgtcta tccatttccc aaccgactac ccattcaagc caccaaagat ctccttcaca
     1441 accaagatat atcatccaaa tatcaatgcc aatggtaaca tctgtctgga catcctaaag
     1501 gatcaatggt ctccagctct aactctatcg aaggtcctat tatccatctg ttctttgtta
     1561 acagacgcta atcctgacga tcctttagta ccagaaatcg ctcatatcta caagactgac
     1621 agacccaagt acgaagctac agccagagaa tggacaaaga aatacgctgt actcgagcac
     1681 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1741 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1801 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
     1861 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1921 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1981 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2041 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2101 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2161 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2221 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2281 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2341 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2401 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2461 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2521 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2581 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2641 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2701 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2761 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2821 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2881 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2941 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     3001 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3061 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3121 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3181 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3241 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3301 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3361 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3601 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3781 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4021 atgctcgtc
//
