LOCUS       sequence-459169-        6438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MYC stability reporter construct of the DNA-binding-domain (residues
            340-439) fused to an N-terminal NLS for transient expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6438)
  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 6438)
  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-NLS-Myc(340-439)-GFP-IRES-mCherry (NLS-DBD)
FEATURES             Location/Qualifiers
     source          1..6438
                     /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             94..114
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             337..366
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             442..1161
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    1169..1741
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1336..1353)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1563..1582
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1750..2460
                     /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(1895..1913)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2490..2624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2514..2533)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2568..2587
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2737..2757)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2737..2755)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2776..2792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2776..2792)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2789..2811)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2800..2816
                     /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(2824..2830,2831..2848,2849..2854))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2869..2890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3007..3024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3178..3766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3258..3277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3937..4728,4729..4797))
                     /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     4560..4579
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4798..4902)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4928..5383)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5065..5086)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5277..5296
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5454..6438
                     /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 atgcccaaga aaaaacgcaa ggtgggcagc
      121 ggctcccgaa aatgcaccag ccccaggtcc tcggacaccg aggagaatgt caagaggcga
      181 acacacaacg tcttggagcg ccagaggagg aacgagctaa aacggagctt ttttgccctg
      241 cgtgaccaga tcccggagtt ggaaaacaat gaaaaggccc ccaaggtagt tatccttaaa
      301 aaagccacag catacatcct gtccgtccaa gcagaggagc aaaagctcat ttctgaagag
      361 gacttgttgc ggaaacgacg agaacagttg aaacacaaac ttgaacagct acggaactct
      421 tgtgcgggag gcggaggcag catggtgagc aagggcgccg agctgttcac cggcatcgtg
      481 cccatcctga tcgagctgaa tggcgatgtg aatggccaca agttcagcgt gagcggcgag
      541 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      601 ctgcctgtgc cctggcccac cctggtgacc accctgagct acggcgtgca gtgcttctca
      661 cgctaccccg atcacatgaa gcagcacgac ttcttcaaga gcgccatgcc tgagggctac
      721 atccaggagc gcaccatctt cttcgaggat gacggcaact acaagtcgcg cgccgaggtg
      781 aagttcgagg gcgataccct ggtgaatcgc atcgagctga ccggcaccga tttcaaggag
      841 gatggcaaca tcctgggcaa taagatggag tacaactaca acgcccacaa tgtgtacatc
      901 atgaccgaca aggccaagaa tggcatcaag gtgaacttca agatccgcca caacatcgag
      961 gatggcagcg tgcagctggc cgaccactac cagcagaata cccccatcgg cgatggccct
     1021 gtgctgctgc ccgataacca ctacctgtcc acccagagcg ccctgtccaa ggaccccaac
     1081 gagaagcgcg atcacatgat ctacttcggc ttcgtgaccg ccgccgccat cacccacggc
     1141 atggatgagc tgtacaagtg atctagagcc cctctccctc ccccccccct aacgttactg
     1201 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     1261 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     1321 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     1381 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     1441 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     1501 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     1561 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     1621 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     1681 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat
     1741 acctccggaa tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1801 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1861 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     1921 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     1981 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     2041 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     2101 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     2161 ggccccgtaa tgcagaagaa aaccatgggc tgggaggcct cctccgagcg gatgtacccc
     2221 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     2281 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     2341 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     2401 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtga
     2461 tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     2521 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2581 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2641 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg
     2701 cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     2761 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2821 acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2881 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2941 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3001 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3061 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3121 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3181 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3241 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3301 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3361 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3421 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3481 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3541 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3601 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3661 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3721 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3781 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3841 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3901 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3961 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4021 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4081 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4141 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4201 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4261 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4321 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4381 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4441 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4501 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4561 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4621 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4681 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4741 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4801 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4861 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4921 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4981 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5041 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5101 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5161 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5221 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5281 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     5341 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     5401 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc
     5461 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
     5521 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
     5581 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
     5641 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
     5701 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
     5761 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
     5821 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
     5881 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
     5941 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
     6001 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
     6061 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
     6121 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
     6181 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
     6241 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
     6301 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
     6361 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
     6421 aatgctcgtt tagggaac
//
