LOCUS       sequence-459167-        6993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MYC stability reporter construct with deletion of the N-terminal 
            region (residues 1-143) and point mutations corresponding to the 
            deletion of degron 1 and 2 for transient expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6993)
  AUTHORS   Mark KG, Kolla S, Aguirre JD, Garshott DM, Schmitt S, Haakonsen DL, 
            Xu C, Kater L, Kempf G, Martinez-Gonzalez B, Akopian D, See SK, 
            Thoma NH, Rape M
  TITLE     Orphan quality control shapes network dynamics and gene expression.
  JOURNAL   Cell. 2023 Aug 3;186(16):3460-3475.e23. doi: 
            10.1016/j.cell.2023.06.015. Epub 2023 Jul 20.
  PUBMED    37478862
REFERENCE   2  (bases 1 to 6993)
  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: 
            "10.1016/j.cell.2023.06.015"; journalName: "Cell"; date: "2023-08-3-
            3"; volume: "186"; issue: "16"; pages: "3460-3475.e23"
COMMENT     Sequence Label: pCS2-Myc(Delta-1-143; 
            Delta-Deg1-Delta-Deg2)-GFP-IRES-mCherry
            ...
FEATURES             Location/Qualifiers
     source          1..6993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             622..648
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             892..921
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             997..1716
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    1724..2296
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1891..1908)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2118..2137
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2305..3015
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2450..2468)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3045..3179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3069..3088)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3123..3142
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3292..3312)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3292..3310)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3331..3347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3331..3347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3344..3366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3355..3371
                     /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(3379..3385,3386..3403,3404..3409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3424..3445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3562..3579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3733..4321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3813..3832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4492..5283,5284..5352))
                     /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     5115..5134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5353..5457)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5483..5938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5620..5641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5832..5851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6009..6993
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcgga tcccgccacc atgctggtct cggagaagct ggcctcctac
      121 caggctgcgc gcaaagacag cggcagcccg aaccccgccc gcggccacag cgtctgctcc
      181 acctccagct tgtacctgca ggatctgagc gccgccgcct cagagtgcat cgacccctcg
      241 gtggtcttcc cctaccctct caacgacagc agctcgccca agtcctgcgc ctcgcaagac
      301 tccagcgcct tctctccgtc ctcggattct ctgctctcct cgacggagtc ctccccgcag
      361 ggcagccccg agcccctggt gctccatgag gagacaccgc ccaccaccag cagcgactct
      421 gaggaggaac aagaagatga ggaagaaatc gatgttgttt ctgtggaaaa gaggcaggct
      481 cctggcaaaa ggtcagagtc tggatcacct tctgctggag gccacagcaa acctcctcac
      541 agcccactgg tcctcaagag gtgccacgtc tccacacatc agcacaacta cgcagcgcct
      601 ccctccactc ggaaggacta tcctgctgcc aagagggtca agttggacag tgtcagagtc
      661 ctgagacaga tcagcaacaa ccgaaaatgc accagcccca ggtcctcgga caccgaggag
      721 aatgtcgagg aggagacaca caacgtcttg gagcgccaga ggaggaacga gctaaaacgg
      781 agcttttttg ccctgcgtga ccagatcccg gagttggaaa acaatgaaaa ggcccccaag
      841 gtagttatcc ttaaaaaagc cacagcagcc gccgcggccg cccaagcaga ggagcaaaag
      901 ctcatttctg aagaggactt gttgcggaaa cgacgagaac agttgaaaca caaacttgaa
      961 caggagcgga actctgaggc gggaggcgga ggcagcatgg tgagcaaggg cgccgagctg
     1021 ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg ccacaagttc
     1081 agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1141 tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct gagctacggc
     1201 gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt caagagcgcc
     1261 atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg caactacaag
     1321 tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga gctgaccggc
     1381 accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa ctacaacgcc
     1441 cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa cttcaagatc
     1501 cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca gaataccccc
     1561 atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca gagcgccctg
     1621 tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt gaccgccgcc
     1681 gccatcaccc acggcatgga tgagctgtac aagtgatcta gagcccctct ccctcccccc
     1741 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     1801 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     1861 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     1921 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     1981 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2041 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2101 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2161 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     2221 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     2281 gaaaaacacg ataatacctc cggaatggtg agcaagggcg aggaggataa catggccatc
     2341 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     2401 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     2461 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     2521 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     2581 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     2641 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     2701 aacttcccct ccgacggccc cgtaatgcag aagaaaacca tgggctggga ggcctcctcc
     2761 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     2821 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     2881 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     2941 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     3001 gagctgtaca agtgatctag aactatagtg agtcgtatta cgtagatcca gacatgataa
     3061 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3121 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3181 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     3241 aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     3301 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3361 tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc
     3421 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3481 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3541 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4141 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4561 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4621 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4681 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4741 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4801 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4861 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4921 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4981 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5041 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5101 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5161 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5221 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5281 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5341 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5401 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5461 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     5521 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5581 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5641 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5701 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5761 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5821 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5881 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     5941 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6001 ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
     6061 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
     6121 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg
     6181 cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga
     6241 ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg
     6301 ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa
     6361 tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag tattccatat
     6421 atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat
     6481 ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg
     6541 gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg
     6601 acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc
     6661 caccattgac gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat
     6721 tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact
     6781 tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt
     6841 caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga
     6901 ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg
     6961 gaggtctata taagcaatgc tcgtttaggg aac
//
