LOCUS       sequence-456428-        7214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Myc-tagged CYLD catalytic inactive (C601A) expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7214)
  AUTHORS   Stegmeier F, Sowa ME, Nalepa G, Gygi SP, Harper JW, Elledge SJ
  TITLE     The tumor suppressor CYLD regulates entry into mitosis.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 May 22. 104(21):8869-74.
  PUBMED    17495026
REFERENCE   2  (bases 1 to 7214)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2007 May 22. 104(21):8869-74."
COMMENT     Sequence Label: Myc-CYLD-C601A
FEATURES             Location/Qualifiers
     source          1..7214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1081..1110
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1120..1149
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1159..1188
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1198..1227
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1237..1266
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1300..1329
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    4244..4378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4268..4287)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4322..4341
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4491..4511)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4491..4509)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4530..4546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4530..4546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4543..4565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4554..4570
                     /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(4578..4584,4585..4602,4603..4608))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4623..4644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4761..4778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4932..5520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5012..5031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5691..6482,6483..6551))
                     /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     6314..6333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6552..6656)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6682..7137)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6819..6840)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7031..7050
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggtggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt taaagctatg
     1081 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
     1141 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
     1201 caaaagctca tttctgaaga ggacttgaat gaaatggagc aaaagctcat ttctgaagag
     1261 gacttgaatg aaatggagag cttgggcgac ctcaccatgg agcaaaagct catttctgaa
     1321 gaggacttga attcaggccg gcccatgagt tcaggcttat ggagccaaga aaaagtcact
     1381 tcaccctact gggaagagcg gattttttac ttgcttcttc aagaatgcag cgttacagac
     1441 aaacaaacac aaaagctcct taaagtaccg aagggaagta taggacagta tattcaagat
     1501 cgttctgtgg ggcattcaag gattccttct gcaaaaggca agaaaaatca gattggatta
     1561 aaaattctag agcaacctca tgcagttctc tttgttgatg aaaaggatgt tgtagagata
     1621 aatgaaaagt tcacagagtt acttttggca attaccaatt gtgaggagag gttcagcctg
     1681 tttaaaaaca gaaacagact aagtaaaggc ctccaaatag acgtgggctg tcctgtgaaa
     1741 gtacagctga gatctgggga agaaaaattt cctggagttg tacgcttcag aggacccctg
     1801 ttagcagaga ggacagtctc cggaatattc tttggagttg aattgctgga agaaggtcgt
     1861 ggtcaaggtt tcactgacgg ggtgtaccaa gggaaacagc tttttcagtg tgatgaagat
     1921 tgtggcgtgt ttgttgcatt ggacaagcta gaactcatag aagatgatga cactgcattg
     1981 gaaagtgatt acgcaggtcc tggggacaca atgcaggtcg aacttcctcc tttggaaata
     2041 aactccagag tttctttgaa ggttggagaa acaatagaat ctggaacagt tatattctgt
     2101 gatgttttgc caggaaaaga aagcttagga tattttgttg gtgtggacat ggataaccct
     2161 attggcaact gggatggaag atttgatgga gtgcagcttt gtagttttgc gtgtgttgaa
     2221 agtacaattc tattgcacat caatgatatc atcccagaga gtgtgacgca ggaaaggagg
     2281 cctcccaaac ttgcctttat gtcaagaggt gttggggaca aaggttcatc cagtcataat
     2341 aaaccaaagg ctacaggatc tacctcagac cctggaaata gaaacagatc tgaattattt
     2401 tataccttaa atgggtcttc tgttgactca caaccacaat ccaaatcaaa aaatacatgg
     2461 tacattgatg aagttgcaga agaccctgca aaatctctta cagagatatc tacagacttt
     2521 gaccgttctt caccaccact ccagcctcct cctgtgaact cactgaccac cgagaacaga
     2581 ttccactctt taccattcag tctcaccaag atgcccaata ccaatggaag tattggccac
     2641 agtccacttt ctctgtcagc ccagtctgta atggaagagc taaacactgc acccgtccaa
     2701 gagagtccac ccttggccat gcctcctggg aactcacatg gtctagaagt gggctcattg
     2761 gctgaagtta aggagaaccc tcctttctat ggggtaatcc gttggatcgg tcagccacca
     2821 ggactgaatg aagtgctcgc tggactggaa ctggaagatg agtgtgcagg ctgtacggat
     2881 ggaaccttca gaggcactcg gtatttcacc tgtgccctga agaaggcgct gtttgtgaaa
     2941 ctgaagagct gcaggcctga ctctaggttt gcatcattgc agccggtttc caatcagatt
     3001 gagcgctgta actctttagc atttggaggc tacttaagtg aagtagtaga agaaaatact
     3061 ccaccaaaaa tggaaaaaga aggcttggag ataatgattg ggaagaagaa aggcatccag
     3121 ggtcattaca attctgctta cttagactca accttattct gcttatttgc ttttagttct
     3181 gttctggaca ctgtgttact tagacccaaa gaaaagaacg atgtagaata ttatagtgaa
     3241 acccaagagc tactgaggac agaaattgtt aatcctctga gaatatatgg atatgtgtgt
     3301 gccacaaaaa ttatgaaact gaggaaaata cttgaaaagg tggaggctgc atcaggattt
     3361 acctctgaag aaaaagatcc tgaggaattc ttgaatattc tgtttcatca tattttaagg
     3421 gtagaacctt tgctaaaaat aagatcagca ggtcaaaagg tacaagattg ttacttctat
     3481 caaattttta tggaaaaaaa tgagaaagtt ggcgttccca caattcagca gttgttagaa
     3541 tggtctttta tcaacagtaa cctgaaattt gcagaggcac catcatgtct gattattcag
     3601 atgcctcgat ttggaaaaga ctttaaacta tttaaaaaaa tttttccttc tctggaatta
     3661 aatataacag atttacttga agacactccc agacagtgcc ggatatgtgg agggcttgca
     3721 atgtatgagt gtagagaatg ctacgacgat ccggacatct cagctggaaa aatcaagcag
     3781 ttttgtaaaa cctgcaacac tcaagtccac cttcatccga agaggctgaa tcataaatat
     3841 aacccagtgt cacttcccaa agacttaccc gactgggact ggagacacgg ctgcatccct
     3901 tgccagaata tggagttatt tgctgttctc tgcatagaaa caagccacta tgttgctttt
     3961 gtgaagtatg ggaaggacga ttctgcctgg ctcttctttg acagcatggc cgatcgggat
     4021 ggtggtcaga atggcttcaa cattcctcaa gtcaccccat gcccagaagt aggagagtac
     4081 ttgaagatgt ctctggaaga cctgcattcc ttggactcca ggagaatcca aggctgtgca
     4141 cgaagactgc tttgtgatgc atatatgtgc atgtaccaga gtccaacaat gagtttgtac
     4201 aaataaggcg cgcctctaga actatagtga gtcgtattac gtagatccag acatgataag
     4261 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4321 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4381 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     4441 attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga
     4501 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4561 ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc
     4621 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4681 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4741 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4801 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4861 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4921 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4981 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5101 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5341 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5401 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5581 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5641 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5701 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5761 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5821 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5881 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5941 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6001 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6061 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6121 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6181 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6241 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6301 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6361 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6421 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6481 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6541 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6601 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6661 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     6721 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     6781 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     6841 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     6901 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     6961 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     7021 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     7081 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     7141 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     7201 tacgccagtc gacc
//
