LOCUS       sequence-452247-        4363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of yeast Rpn10 with the UIM deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4363)
  AUTHORS   Jonsson E, Htet ZM, Bard JAM, Dong KC, Martin A
  TITLE     Ubiquitin modulates 26S proteasome conformational dynamics and 
            promotes substrate degradation.
  JOURNAL   Sci Adv. 2022 Dec 23;8(51):eadd9520. doi: 10.1126/sciadv.add9520. 
            Epub 2022 Dec 23.
  PUBMED    36563145
REFERENCE   2  (bases 1 to 4363)
  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"; journalName: "Sci Adv."; 
            date: "2022-12-23"; volume: "8(51):eadd9520. doi"; pages: " 
            10.1126/sciadv.add9520. Epub 2022 Dec 23"
COMMENT     Sequence Label: pAM315 pACYC-His6-Rpn10[Delta-UIM]
FEATURES             Location/Qualifiers
     source          1..4363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..20)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(691..1773)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1735..1754
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1774..1851)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1897..1916
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1897..1915
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1916..1940
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             1990..2007
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2632..2676
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2714..2732)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2728..2775
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2996..3655)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3570..3589
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3656..3758)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4150..4167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4284..4363,1..466))
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
ORIGIN
        1 aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc
       61 ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc
      121 agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc
      181 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa
      241 gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc
      301 cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc
      361 tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt
      421 ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat
      481 taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga
      541 agtcagcccc atacgatata agttgtaatt ctcatgttag tcatgccccg cgcccaccgg
      601 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
      661 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      721 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      781 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
      841 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
      901 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
      961 cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     1021 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     1081 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     1141 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     1201 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     1261 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     1321 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     1381 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     1441 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     1501 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     1561 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     1621 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     1681 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     1741 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     1801 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     1861 gacgctctcc cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat
     1921 tgtgagcgga taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat
     1981 accatgggac atcaccatca ccatcacggc tcagtattgg aagctacagt gttagtgatt
     2041 gataattcag agtactctcg caatggcgac tttcctagga caaggtttga agcccagatc
     2101 gactcggtag agttcatatt tcaagccaag agaaacagca atcctgagaa tacagtgggt
     2161 cttatctctg gtgctggcgc caaccccagg gtgttatcta cgtttaccgc cgagtttggg
     2221 aagattcttg ctggactaca cgacacgcag atcgagggta agctgcatat ggccactgcg
     2281 ttgcagatcg ctcagctgac tttgaagcat cgccagaata aggtccaaca tcaaaggatt
     2341 gtggcatttg tgtgtagccc aataagtgat tctcgagacg aattgatcag attggcaaaa
     2401 acactgaaaa agaataatgt tgccgtggac atcatcaatt ttggagagat tgaacagaac
     2461 acggagcttt tggatgagtt catagctgca gtgaacaacc ctcaagaaga aactagtcat
     2521 ttgcttactg tgacgcctgg ccccagactg ctgtacgaga acatcgcatc ttcacccata
     2581 attctcgaag aaggatcctc ctaaaagctt gcggccgcac tcgagtctgg taaagaaacc
     2641 gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc
     2701 taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2761 ttgaggggtt ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg
     2821 gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct
     2881 gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac
     2941 ttattcaggc gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg
     3001 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     3061 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     3121 gcgtataata tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     3181 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
     3241 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     3301 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     3361 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     3421 ctttcattgc catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa
     3481 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     3541 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     3601 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     3661 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     3721 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt
     3781 tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat
     3841 cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga
     3901 tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc
     3961 tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc
     4021 tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
     4081 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
     4141 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     4201 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     4261 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
     4321 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tat
//
