LOCUS       sequence-455233-        7417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS305 His-HRV3C-Rpn5 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7417)
  AUTHORS   Greene ER, Goodall EA, de la Pena AH, Matyskiela ME, Lander GC, 
            Martin A
  TITLE     Specific lid-base contacts in the 26s proteasome control the 
            conformational switching required for substrate degradation.
  JOURNAL   Elife. 2019 Nov 28;8:e49806. doi: 10.7554/eLife.49806.
  PUBMED    31778111
REFERENCE   2  (bases 1 to 7417)
  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: "Elife."; 
            date: "2019-11-28"; volume: "8:e49806. doi"; pages: " 
            10.7554/eLife.49806"
COMMENT     Sequence Label: pAM198
FEATURES             Location/Qualifiers
     source          1..7417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(162..569)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(870..1325)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1007..1028)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1219..1238
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1451..1473
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1465..1482
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1466..1482
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1489..1508
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1489..1507
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2962..2985)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(2995..3024)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(3560..3578)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3599..3615)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3599..3615)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3612..3634)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3623..3639
                     /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)."
     promoter        complement(join(3647..3653,3654..3671,3672..3677))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3692..3713
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3830..3847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4001..4589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4081..4100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4760..5551,5552..5620))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5383..5402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5621..5725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5793..5811
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5849..5871)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5971..5990
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(join(6484..7417,1..161))
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 gtagcatcga tagcagcacc accaattaaa tgattttcga aatcgaactt gacattggaa
       61 cgaacatcag aaatagcttt aagaacctta atggcttcgg ctgtgatttc ttgaccaacg
      121 tggtcacctg gcaaaacgac gatcttctta ggggcagaca ttagaatggt atatccttga
      181 aatatatata tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat
      241 tgctaaccac ctattggaaa aaacaatagg tccttaaata atattgtcaa cttcaagtat
      301 tgtgatgcaa gcatttagtc atgaacgctt ctctattcta tatgaaaagc cggttccggc
      361 gctctcacct ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga
      421 cctctgaaat taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct
      481 gtgtgttctc gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca
      541 tcttacgata cctgagtatt cccacagtta actgcggtca agatatttct tgaatcaggc
      601 gccttagacc gctcggccaa acaaccaatt acttgttgag aaatagagta taattatcct
      661 ataaatataa cgtttttgaa cacacatgaa caaggaagta caggacaatt gattttgaag
      721 agaatgtgga ttttgatgta attgttggga ttccattttt aataaggcaa taatattagg
      781 tatgtagata tactagaagt tctcctcgac cggtcgatat gcggtgtgaa ataccgcaca
      841 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
      901 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
      961 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1021 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1081 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     1141 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1201 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     1261 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     1321 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     1381 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     1441 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     1501 ctatagggcg aattggagct cataatttta tctcacgtac gtaaatacac atttcgatta
     1561 gtgacagatt taaaatcaaa accggatctg agataatccg acagttactc gaaaatctct
     1621 tcatttagct tgcagaccgt gcatgatttc ctcttttgta attaaatggc ctattgtttc
     1681 tatatgttct aatagttcgt caacattatg tgaccattcg ttcaataatt ggcttgagtt
     1741 ttttggtttt tcaaaattca cgattttggc tgggcgatta actttagcgt atatgatgcc
     1801 ctggtttacc aaatcactga tgtatgtttc cgtctggctc tccgttaggt ccagcaattc
     1861 attcaatctt agtaaagtaa ttctggaata gtattcggaa atgactctta aattgtgctc
     1921 gatgaccctt ttttgtaaat cttcccagtg atgcttatta gcttctccac caaatgccaa
     1981 atcatcctca tttaagacgg gctcataggt tttttgaaca attggccatc tcatcaactc
     2041 attcgtagta aacaatttta ctaaagattc ttggctttct aattttttca ggttgttatc
     2101 attctggatt ttgtgaatta aatcattttg taaattgccg taaggtgaaa ggactaagaa
     2161 atatacaatg tgcgataaaa caggtttcca cttagcctca tctgatttaa tggcgtctgt
     2221 ttgataaatt tcttgcagat actgcgcaac ttctaggtat tctctcttgt gcaaactaat
     2281 ttttaccaga agattataat attctagctt caatgactcg tattttggat ttttaaaagt
     2341 ttttttcaga atttttcttg aaagcaccgt ggcttgggaa taatcacctt ttaatatact
     2401 caattccatt tgctctaata taaactgaat tttctcagac atttccatgg agccataggt
     2461 ctcaacctgt aactcacaca agatgtctgc agcttcatca atcttaccct cttctttctt
     2521 aatttctacc aaatctttgg tgaccctagc tctttccact tctacagctg ctgcggccgc
     2581 tgcagccacc ctgatagttt caatgacact aattctggtg tttaaatcca aagatttcga
     2641 gcttttcaaa tattccataa ccttttgtat catatactga attgacaatt tcaactgacc
     2701 atgctttttt gagagtagag tcaattgctc atttaggtcg tcccacttat tccttgatgc
     2761 tagcagatct acaatcttgg ccaaaacttc tttcgaggag gccagatctg aagcttgtct
     2821 ggttttcttc tccaacacta acagttggtc taaagcagag ttacaatcat tttgagcgag
     2881 cgaatcgatc ttaggaaact cttccttcaa aatttggcta taatccttgt cagccttaat
     2941 tggtgcatct cttgacatac ctggaccttg aaataaaact tccaaaccgc ttccgtgatg
     3001 gtgatggtga tggtgatggt gatgcatctt tctctacgtt tactctagca acaaaaatgg
     3061 tactgagccc agcagtatat tcctctactc tcttaattca agattatctc tgatgtagtg
     3121 tcctgttatg gtttttcctt agattcccct ttgcattttg ccaccgaaaa gtcaaaaaga
     3181 tcacatacaa gacgggtaat attggggcga tgagccgtac agaacgagtg aaaaatttta
     3241 ggatttatta ggaataataa tgataattta gagatgagta tttagcgaga agcatctgaa
     3301 ggttgcacta attattactg taaatagtac agttgagaag caaatacaag ggaagatcaa
     3361 atacatggcc ggttcacaac ttaagaattt aaaggccgcc ttgaaggccc gtggactcac
     3421 aggtcaaaca aacgtcaaaa gtaagaataa aaaaaattcc aaaagacaag cgaaggagta
     3481 tgatcgtgaa gaaaaaaaaa aggccattgc tgaaatcaga gaagagtctc gagggggggc
     3541 ccggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg taatcatggt
     3601 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     3661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3841 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3901 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3961 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4021 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4081 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4141 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4201 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4261 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4321 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4381 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4441 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4501 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4561 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4621 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4681 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4741 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4861 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4921 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5281 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5641 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5701 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5761 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5821 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5881 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5941 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     6001 accatatcga ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt
     6061 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc
     6121 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa
     6181 attaggaatc atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct
     6241 ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta
     6301 tcaatttgct tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg
     6361 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc
     6421 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc
     6481 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact
     6541 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc
     6601 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga
     6661 caagatagtg gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca
     6721 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg
     6781 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat
     6841 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc
     6901 agaatcaatc aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc
     6961 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat
     7021 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac
     7081 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct
     7141 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc
     7201 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct
     7261 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac
     7321 actaccggta ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt
     7381 ggaggcttcc agcgcctcat ctggaagtgg aacacct
//
