LOCUS       sequence-459172-        6432 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 NES for transient expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6432)
  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 6432)
  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-NES-Myc(340-439)-GFP-IRES-mCherry (NES-DBD)
FEATURES             Location/Qualifiers
     source          1..6432
                     /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..120
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             331..360
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             436..1155
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    1163..1735
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1330..1347)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1557..1576
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1744..2454
                     /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(1889..1907)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2484..2618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2508..2527)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2562..2581
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2731..2751)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2731..2749)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2770..2786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2770..2786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2783..2805)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2794..2810
                     /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(2818..2824,2825..2842,2843..2848))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2863..2884
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3001..3018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3172..3760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3252..3271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3931..4722,4723..4791))
                     /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     4554..4573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4792..4896)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4922..5377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5059..5080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5271..5290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5448..6432
                     /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 atgctgcccc ccctggagcg cctgaccctg
      121 cgaaaatgca ccagccccag gtcctcggac accgaggaga atgtcaagag gcgaacacac
      181 aacgtcttgg agcgccagag gaggaacgag ctaaaacgga gcttttttgc cctgcgtgac
      241 cagatcccgg agttggaaaa caatgaaaag gcccccaagg tagttatcct taaaaaagcc
      301 acagcataca tcctgtccgt ccaagcagag gagcaaaagc tcatttctga agaggacttg
      361 ttgcggaaac gacgagaaca gttgaaacac aaacttgaac agctacggaa ctcttgtgcg
      421 ggaggcggag gcagcatggt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc
      481 ctgatcgagc tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag
      541 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct
      601 gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac
      661 cccgatcaca tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag
      721 gagcgcacca tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc
      781 gagggcgata ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc
      841 aacatcctgg gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc
      901 gacaaggcca agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc
      961 agcgtgcagc tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg
     1021 ctgcccgata accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag
     1081 cgcgatcaca tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat
     1141 gagctgtaca agtgatctag agcccctctc cctccccccc ccctaacgtt actggccgaa
     1201 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     1261 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     1321 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     1381 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     1441 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     1501 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     1561 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     1621 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     1681 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga taatacctcc
     1741 ggaatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     1801 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     1861 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1921 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1981 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2041 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2101 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2161 gtaatgcaga agaaaaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2221 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2281 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2341 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2401 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtgatctaga
     2461 actatagtga gtcgtattac gtagatccag acatgataag atacattgat gagtttggac
     2521 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     2581 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     2641 ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa
     2701 tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     2761 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2821 catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2881 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2941 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3061 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4921 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4981 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     5041 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     5101 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     5161 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     5221 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     5281 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     5341 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     5401 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccgccaat
     5461 tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac ctgtgccaat
     5521 tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac cccagccaat
     5581 tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacctggca ctgtgccaac
     5641 tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc cctgtgccaa
     5701 gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct atatgccagg
     5761 atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg gccaggttca
     5821 atactatgta ttggccctat gccatatagt attccatata tgggttttcc tattgacgta
     5881 gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta ataccgccca
     5941 tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat tgacgtcata
     6001 tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac ggtaaatggc
     6061 ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg tcaatgggat
     6121 ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg acggtaaatg
     6181 gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca ttactattgg
     6241 aagtacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta aatggcccgc
     6301 gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac gcaaatgggc
     6361 gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat aagcaatgct
     6421 cgtttaggga ac
//
