LOCUS       sequence-459171-        6738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MYC stability reporter construct with deletion of the N-terminal 
            region (residues 1-143) and DNA-binding-domain (residues 340-439) 
            for transient expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6738)
  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 6738)
  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-340-439)-GFP-IRES-mCherry
            (Delta...
FEATURES             Location/Qualifiers
     source          1..6738
                     /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             742..1461
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    1469..2041
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1636..1653)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1863..1882
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2050..2760
                     /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(2195..2213)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2790..2924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2814..2833)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2868..2887
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3037..3057)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3037..3055)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3076..3092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3076..3092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3089..3111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3100..3116
                     /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(3124..3130,3131..3148,3149..3154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3169..3190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3307..3324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3478..4066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3558..3577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4237..5028,5029..5097))
                     /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     4860..4879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5098..5202)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5228..5683)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5365..5386)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5577..5596
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5754..6738
                     /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 aatgtcggag gcggaggcag catggtgagc aagggcgccg agctgttcac cggcatcgtg
      781 cccatcctga tcgagctgaa tggcgatgtg aatggccaca agttcagcgt gagcggcgag
      841 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      901 ctgcctgtgc cctggcccac cctggtgacc accctgagct acggcgtgca gtgcttctca
      961 cgctaccccg atcacatgaa gcagcacgac ttcttcaaga gcgccatgcc tgagggctac
     1021 atccaggagc gcaccatctt cttcgaggat gacggcaact acaagtcgcg cgccgaggtg
     1081 aagttcgagg gcgataccct ggtgaatcgc atcgagctga ccggcaccga tttcaaggag
     1141 gatggcaaca tcctgggcaa taagatggag tacaactaca acgcccacaa tgtgtacatc
     1201 atgaccgaca aggccaagaa tggcatcaag gtgaacttca agatccgcca caacatcgag
     1261 gatggcagcg tgcagctggc cgaccactac cagcagaata cccccatcgg cgatggccct
     1321 gtgctgctgc ccgataacca ctacctgtcc acccagagcg ccctgtccaa ggaccccaac
     1381 gagaagcgcg atcacatgat ctacttcggc ttcgtgaccg ccgccgccat cacccacggc
     1441 atggatgagc tgtacaagtg atctagagcc cctctccctc ccccccccct aacgttactg
     1501 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     1561 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     1621 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     1681 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     1741 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     1801 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     1861 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     1921 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     1981 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat
     2041 acctccggaa tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     2101 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     2161 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     2221 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     2281 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     2341 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     2401 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     2461 ggccccgtaa tgcagaagaa aaccatgggc tgggaggcct cctccgagcg gatgtacccc
     2521 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     2581 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     2641 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     2701 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtga
     2761 tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     2821 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2881 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2941 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg
     3001 cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     3061 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3121 acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3181 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3241 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3301 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3361 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3421 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3481 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3541 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3601 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3661 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3721 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3781 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3841 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3901 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3961 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4081 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4141 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4201 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4261 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4321 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4381 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4441 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4501 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4561 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4621 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4681 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4741 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4801 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4861 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4921 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4981 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5041 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5101 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5161 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5221 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5281 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5341 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5401 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5461 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5521 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5581 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5641 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     5701 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc
     5761 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
     5821 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
     5881 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
     5941 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
     6001 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
     6061 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
     6121 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
     6181 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
     6241 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
     6301 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
     6361 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
     6421 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
     6481 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
     6541 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
     6601 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
     6661 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
     6721 aatgctcgtt tagggaac
//
