LOCUS       sequence-458628-        7419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MYC stability reporter construct (expressing c-myc 2) for transient 
            expression in mammalian cells. Stability of GFP-fusion protein can 
            be assed by flow cytometry by normalizing to mCherry expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7419)
  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 7419)
  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-GFP-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..7419
                     /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             91..1407
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     CDS             1423..2142
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    2150..2722
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2317..2334)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2544..2563
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2731..3441
                     /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(2876..2894)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3471..3605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3495..3514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3549..3568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3718..3738)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3718..3736)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3757..3773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3757..3773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3770..3792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3781..3797
                     /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(3805..3811,3812..3829,3830..3835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3850..3871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3988..4005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4159..4747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4239..4258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4918..5709,5710..5778))
                     /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     5541..5560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5779..5883)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5909..6364)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6046..6067)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6258..6277
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6435..7419
                     /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 atgcccctca acgttagctt caccaacagg
      121 aactatgacc tcgactacga ctcggtgcag ccgtatttct actgcgacga ggaggagaac
      181 ttctaccagc agcagcagca gagcgagctg cagcccccgg cgcccagcga ggatatctgg
      241 aagaaattcg agctgctgcc caccccgccc ctgtccccta gccgccgctc cgggctctgc
      301 tcgccctcct acgttgcggt cacacccttc tcccttcggg gagacaacga cggcggtggc
      361 gggagcttct ccacggccga ccagctggag atggtgaccg agctgctggg aggagacatg
      421 gtgaaccaga gtttcatctg cgacccggac gacgagacct tcatcaaaaa catcatcatc
      481 caggactgta tgtggagcgg cttctcggcc gccgccaagc tcgtctcaga gaagctggcc
      541 tcctaccagg ctgcgcgcaa agacagcggc agcccgaacc ccgcccgcgg ccacagcgtc
      601 tgctccacct ccagcttgta cctgcaggat ctgagcgccg ccgcctcaga gtgcatcgac
      661 ccctcggtgg tcttccccta ccctctcaac gacagcagct cgcccaagtc ctgcgcctcg
      721 caagactcca gcgccttctc tccgtcctcg gattctctgc tctcctcgac ggagtcctcc
      781 ccgcagggca gccccgagcc cctggtgctc catgaggaga caccgcccac caccagcagc
      841 gactctgagg aggaacaaga agatgaggaa gaaatcgatg ttgtttctgt ggaaaagagg
      901 caggctcctg gcaaaaggtc agagtctgga tcaccttctg ctggaggcca cagcaaacct
      961 cctcacagcc cactggtcct caagaggtgc cacgtctcca cacatcagca caactacgca
     1021 gcgcctccct ccactcggaa ggactatcct gctgccaaga gggtcaagtt ggacagtgtc
     1081 agagtcctga gacagatcag caacaaccga aaatgcacca gccccaggtc ctcggacacc
     1141 gaggagaatg tcaagaggcg aacacacaac gtcttggagc gccagaggag gaacgagcta
     1201 aaacggagct tttttgccct gcgtgaccag atcccggagt tggaaaacaa tgaaaaggcc
     1261 cccaaggtag ttatccttaa aaaagccaca gcatacatcc tgtccgtcca agcagaggag
     1321 caaaagctca tttctgaaga ggacttgttg cggaaacgac gagaacagtt gaaacacaaa
     1381 cttgaacagc tacggaactc ttgtgcggga ggcggaggca gcatggtgag caagggcgcc
     1441 gagctgttca ccggcatcgt gcccatcctg atcgagctga atggcgatgt gaatggccac
     1501 aagttcagcg tgagcggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1561 ttcatctgca ccaccggcaa gctgcctgtg ccctggccca ccctggtgac caccctgagc
     1621 tacggcgtgc agtgcttctc acgctacccc gatcacatga agcagcacga cttcttcaag
     1681 agcgccatgc ctgagggcta catccaggag cgcaccatct tcttcgagga tgacggcaac
     1741 tacaagtcgc gcgccgaggt gaagttcgag ggcgataccc tggtgaatcg catcgagctg
     1801 accggcaccg atttcaagga ggatggcaac atcctgggca ataagatgga gtacaactac
     1861 aacgcccaca atgtgtacat catgaccgac aaggccaaga atggcatcaa ggtgaacttc
     1921 aagatccgcc acaacatcga ggatggcagc gtgcagctgg ccgaccacta ccagcagaat
     1981 acccccatcg gcgatggccc tgtgctgctg cccgataacc actacctgtc cacccagagc
     2041 gccctgtcca aggaccccaa cgagaagcgc gatcacatga tctacttcgg cttcgtgacc
     2101 gccgccgcca tcacccacgg catggatgag ctgtacaagt gatctagagc ccctctccct
     2161 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     2221 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     2281 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     2341 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     2401 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     2461 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     2521 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     2581 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     2641 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     2701 tcctttgaaa aacacgataa tacctccgga atggtgagca agggcgagga ggataacatg
     2761 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     2821 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2881 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     2941 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     3001 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     3061 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     3121 ggcaccaact tcccctccga cggccccgta atgcagaaga aaaccatggg ctgggaggcc
     3181 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     3241 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     3301 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     3361 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     3421 atggacgagc tgtacaagtg atctagaact atagtgagtc gtattacgta gatccagaca
     3481 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     3541 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3601 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     3661 ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
     3721 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3781 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
     3841 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3901 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3961 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4021 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4081 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4561 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4741 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4801 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4861 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4921 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4981 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5041 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5101 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5161 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5221 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5281 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5341 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5401 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5461 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5521 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5581 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5641 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5701 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5761 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5821 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5881 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     5941 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     6001 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     6061 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     6121 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     6181 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     6241 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     6301 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     6361 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6421 tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca
     6481 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     6541 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     6601 atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     6661 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     6721 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     6781 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     6841 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     6901 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     6961 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     7021 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     7081 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     7141 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     7201 gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat
     7261 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     7321 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     7381 taatgggagg tctatataag caatgctcgt ttagggaac
//
