LOCUS       sequence-9532-ex        6755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6755)
  AUTHORS   Salomons FA, Menendez-Benito V, Bottcher C, McCray BA, Taylor JP, 
            Dantuma NP
  TITLE     Selective accumulation of aggregation-prone proteasome substrates in
            response to proteotoxic stress.
  JOURNAL   Mol Cell Biol. 2009 Apr;29(7):1774-85. doi: 10.1128/MCB.01485-08. 
            Epub 2009 Jan 21.
  PUBMED    19158272
REFERENCE   2  (bases 1 to 6755)
  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: 
            "10.1128/MCB.01485-08"; journalName: "Mol Cell Biol"; date: 
            "2009-04"; volume: "29"; issue: "7"; pages: "1774-85"
COMMENT     Sequence Label: Ub[9]-mRFP[1]
FEATURES             Location/Qualifiers
     source          1..6755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..261
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             262..489
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             718..945
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             946..1173
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             946..1173
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1174..1401
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1174..1401
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1402..1629
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1402..1629
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1630..1857
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1630..1857
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1858..2085
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             2122..2799
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2122..2799
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2252..2270)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2520..2539
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2714..2737
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2921..3042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2958..2977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3012..3031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3049..3504)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3186..3207)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3398..3417
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3531..3635
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3637..3994
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3637..3828
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3660..3680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3845..3980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4029..4823
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4029..4823
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4083..4102)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4693..4712
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5011..5030)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5055..5102
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5431..6019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5920..5939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6194..6497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6498..6701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6652..6672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 agctttgtgg atcgctgtga tcgtcacttg acaatgcaga tcttcgtgaa gactctgact
       61 ggtaagacca tcaccctcga ggttgagccc agtgacacca tcgagaatgt caaggcaaag
      121 atccaagata aggaaggcat ccctcctgac cagcagaggc tgatctttgc tggaaaacag
      181 ctggaagatg ggcgcaccct gtctgactac aacatccaga aagagtccac cctgcacctg
      241 gtgctccgtc tcagaggtgg gatgcaaatc ttcgtgaaga cactcactgg caagaccatc
      301 acccttgagg tcgagcccag tgacaccatc gagaacgtca aagcaaagat ccaggacaag
      361 gaaggcattc ctcctgacca gcagaggttg atctttgccg gaaagcagct ggaagatggg
      421 cgcaccctgt ctgactacaa catccagaaa gagtctaccc tgcacctggt gctccgtctc
      481 agaggtggga tgcagatctt cgtgaagacc ctgactggta agaccatcac cctcgaggtg
      541 gagcccagtg acaccatcga gaatgtcaag gcaaagatcc aagataagga aggcattcct
      601 tctgatcagc agaggttgat ctttgccgga aaacagctgg aagatggtcg taccctgtct
      661 gactacaaca tccagaaaga gtccaccttg cacctggtac tccgtctcag aggtgggatg
      721 caaatcttcg tgaagacact cactggcaag accatcaccc ttgaggtcga gcccagtgac
      781 actatcgaga acgtcaaagc aaagatccaa gacaaggaag gcattcctcc tgaccagcag
      841 aggttgatct ttgccggaaa gcagctggaa gatgggcgca ccctgtctga ctacaacatc
      901 cagaaagagt ctaccctgca cctggtgctc cgtctcagag gtgggatgca gatcttcgtg
      961 aagaccctga ctggtaagac catcactctc gaagtggagc cgagtgacac cattgagaat
     1021 gtcaaggcaa agatccaaga caaggaaggc atccctcctg accagcagag gttgatcttt
     1081 gccggaaaac agctggaaga tggtcgtacc ctgtctgact acaacatcca gaaagagtcc
     1141 accttgcacc tggtgctccg tctcagaggt gggatgcaga tcttcgtgaa gaccctgact
     1201 ggtaagacca tcactctcga ggtggagccg agtgacacca ttgagaatgt caaggcaaag
     1261 atccaagaca aggaaggcat ccctcctgac cagcagaggt tgatctttgc tgggaaacag
     1321 ctggaagatg gacgcaccct gtctgactac aacatccaga aagagtccac cctgcacctg
     1381 gtgctccgtc ttagaggtgg gatgcagatc ttcgtgaaga ccctgactgg taagaccatc
     1441 actctcgaag tggagccgag tgacaccatt gagaatgtca aggcaaagat ccaagacaag
     1501 gaaggcatcc ctcctgacca gcagaggttg atctttgctg ggaaacagct ggaagatgga
     1561 cgcaccctgt ctgactacaa catccagaaa gagtccaccc tgcacctggt gctccgtctt
     1621 agaggtggga tgcagatctt cgtgaagacc ctgactggta agaccatcac tctcgaagta
     1681 gagccgagtg acaccattga gaatgtcaag gcaaagatcc aagacaagga aggcatccct
     1741 cctgaccagc agaggttgat ctttgctggg aaacagctgg aagatggacg caccctgtct
     1801 gactacaaca tccagaaaga gtccaccctg cacctggtgc tccgtctcag aggtgggatg
     1861 cagatcttcg tgaagaccct gactggtaag accatcaccc tcgaggtgga gcccagtgac
     1921 accatcgaga atgtcaaggc aaagatccaa gataaggaag gcatccctcc tgatcagcag
     1981 aggttgatct ttgctgggaa acagctggaa gatggacgca ccctgtctga ctacaacatc
     2041 cagaaagagt ccactctgca cttggtcctg cgcttgaggg ggggtgtcgg agtttcccct
     2101 tttaaggttt caacggatcc gatggcctcc tccgaggacg tcatcaagga gttcatgcgc
     2161 ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     2221 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     2281 ctgcccttcg cctgggacat cctgtcccct cagttccagt acggctccaa ggcctacgtg
     2341 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     2401 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2461 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2521 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcggat gtaccccgag
     2581 gacggcgccc tgaagggcga gatcaagatg aggctgaagc tgaaggacgg cggccactac
     2641 gacgccgagg tcaagaccac ctacatggcc aagaagcccg tgcagctgcc cggcgcctac
     2701 aagaccgaca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2761 tacgagcgcg ccgagggccg ccactccacc ggcgcctaag cggccgcgac tctagatcat
     2821 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2881 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2941 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3001 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3061 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3121 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3181 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3241 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3301 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3361 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3421 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3481 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3541 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3601 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3661 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3721 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3841 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3901 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3961 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4021 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4081 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4141 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4201 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4261 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4321 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4381 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4441 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4501 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4561 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4621 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4681 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4741 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4801 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4861 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4921 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4981 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5041 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5101 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5161 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5221 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5281 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5341 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5401 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5461 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5521 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     5581 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5641 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5701 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5761 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5821 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5881 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5941 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6001 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6061 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6121 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     6181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     6241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     6301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     6361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     6421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     6481 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     6541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     6601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     6661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     6721 tccgctagcg ctaccggact cagatctcga gctca
//
