LOCUS       sequence-219908-        7028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7028)
  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 7028)
  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: HA-CYLD-WT
FEATURES             Location/Qualifiers
     source          1..7028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..980
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1015..1033
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1015..1032
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1071..1151
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    4060..4194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4084..4103)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4138..4157
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4307..4327)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4307..4325)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4346..4362)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4346..4362)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4359..4381)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4370..4386
                     /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(4394..4400,4401..4418,4419..4424))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4439..4460
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4577..4594)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4748..5336)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4828..4847)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5507..6298,6299..6367))
                     /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     6130..6149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6368..6472)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6498..6953)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6635..6656)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6847..6866
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccgccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
       61 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
      121 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact
      181 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
      241 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
      301 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
      361 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
      421 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
      481 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
      541 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
      601 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
      661 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
      721 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
      781 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
      841 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
      901 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
      961 gcaatgctcg tttagggaac cgccattctg cctggggacg tcggagcaag cttgatttag
     1021 gtgacactat agaatacaag ctacttgttc tttttgcagg atccaccatg tacccatacg
     1081 atgttccaga ttacgcttat ccttatgacg tacctgacta tgcataccct tatgatgtac
     1141 cagactacgc tggccggccc atgagttcag gcttatggag ccaagaaaaa gtcacttcac
     1201 cctactggga agagcggatt ttttacttgc ttcttcaaga atgcagcgtt acagacaaac
     1261 aaacacaaaa gctccttaaa gtaccgaagg gaagtatagg acagtatatt caagatcgtt
     1321 ctgtggggca ttcaaggatt ccttctgcaa aaggcaagaa aaatcagatt ggattaaaaa
     1381 ttctagagca acctcatgca gttctctttg ttgatgaaaa ggatgttgta gagataaatg
     1441 aaaagttcac agagttactt ttggcaatta ccaattgtga ggagaggttc agcctgttta
     1501 aaaacagaaa cagactaagt aaaggcctcc aaatagacgt gggctgtcct gtgaaagtac
     1561 agctgagatc tggggaagaa aaatttcctg gagttgtacg cttcagagga cccctgttag
     1621 cagagaggac agtctccgga atattctttg gagttgaatt gctggaagaa ggtcgtggtc
     1681 aaggtttcac tgacggggtg taccaaggga aacagctttt tcagtgtgat gaagattgtg
     1741 gcgtgtttgt tgcattggac aagctagaac tcatagaaga tgatgacact gcattggaaa
     1801 gtgattacgc aggtcctggg gacacaatgc aggtcgaact tcctcctttg gaaataaact
     1861 ccagagtttc tttgaaggtt ggagaaacaa tagaatctgg aacagttata ttctgtgatg
     1921 ttttgccagg aaaagaaagc ttaggatatt ttgttggtgt ggacatggat aaccctattg
     1981 gcaactggga tggaagattt gatggagtgc agctttgtag ttttgcgtgt gttgaaagta
     2041 caattctatt gcacatcaat gatatcatcc cagagagtgt gacgcaggaa aggaggcctc
     2101 ccaaacttgc ctttatgtca agaggtgttg gggacaaagg ttcatccagt cataataaac
     2161 caaaggctac aggatctacc tcagaccctg gaaatagaaa cagatctgaa ttattttata
     2221 ccttaaatgg gtcttctgtt gactcacaac cacaatccaa atcaaaaaat acatggtaca
     2281 ttgatgaagt tgcagaagac cctgcaaaat ctcttacaga gatatctaca gactttgacc
     2341 gttcttcacc accactccag cctcctcctg tgaactcact gaccaccgag aacagattcc
     2401 actctttacc attcagtctc accaagatgc ccaataccaa tggaagtatt ggccacagtc
     2461 cactttctct gtcagcccag tctgtaatgg aagagctaaa cactgcaccc gtccaagaga
     2521 gtccaccctt ggccatgcct cctgggaact cacatggtct agaagtgggc tcattggctg
     2581 aagttaagga gaaccctcct ttctatgggg taatccgttg gatcggtcag ccaccaggac
     2641 tgaatgaagt gctcgctgga ctggaactgg aagatgagtg tgcaggctgt acggatggaa
     2701 ccttcagagg cactcggtat ttcacctgtg ccctgaagaa ggcgctgttt gtgaaactga
     2761 agagctgcag gcctgactct aggtttgcat cattgcagcc ggtttccaat cagattgagc
     2821 gctgtaactc tttagcattt ggaggctact taagtgaagt agtagaagaa aatactccac
     2881 caaaaatgga aaaagaaggc ttggagataa tgattgggaa gaagaaaggc atccagggtc
     2941 attacaattc ttgttactta gactcaacct tattctgctt atttgctttt agttctgttc
     3001 tggacactgt gttacttaga cccaaagaaa agaacgatgt agaatattat agtgaaaccc
     3061 aagagctact gaggacagaa attgttaatc ctctgagaat atatggatat gtgtgtgcca
     3121 caaaaattat gaaactgagg aaaatacttg aaaaggtgga ggctgcatca ggatttacct
     3181 ctgaagaaaa agatcctgag gaattcttga atattctgtt tcatcatatt ttaagggtag
     3241 aacctttgct aaaaataaga tcagcaggtc aaaaggtaca agattgttac ttctatcaaa
     3301 tttttatgga aaaaaatgag aaagttggcg ttcccacaat tcagcagttg ttagaatggt
     3361 cttttatcaa cagtaacctg aaatttgcag aggcaccatc atgtctgatt attcagatgc
     3421 ctcgatttgg aaaagacttt aaactattta aaaaaatttt tccttctctg gaattaaata
     3481 taacagattt acttgaagac actcccagac agtgccggat atgtggaggg cttgcaatgt
     3541 atgagtgtag agaatgctac gacgatccgg acatctcagc tggaaaaatc aagcagtttt
     3601 gtaaaacctg caacactcaa gtccaccttc atccgaagag gctgaatcat aaatataacc
     3661 cagtgtcact tcccaaagac ttacccgact gggactggag acacggctgc atcccttgcc
     3721 agaatatgga gttatttgct gttctctgca tagaaacaag ccactatgtt gcttttgtga
     3781 agtatgggaa ggacgattct gcctggctct tctttgacag catggccgat cgggatggtg
     3841 gtcagaatgg cttcaacatt cctcaagtca ccccatgccc agaagtagga gagtacttga
     3901 agatgtctct ggaagacctg cattccttgg actccaggag aatccaaggc tgtgcacgaa
     3961 gactgctttg tgatgcatat atgtgcatgt accagagtcc aacaatgagt ttgtacaaat
     4021 aaggcgcgcc tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac
     4081 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4141 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4201 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc
     4261 gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt
     4321 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4381 tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat
     4441 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4501 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4561 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4621 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4681 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4741 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4801 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4861 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4921 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4981 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5041 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5101 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5161 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5221 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5281 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5341 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5401 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5461 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5521 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5581 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5641 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5701 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5761 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5821 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5881 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5941 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6001 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6061 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6121 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6181 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6241 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6301 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6361 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6421 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6481 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     6541 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     6601 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     6661 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     6721 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     6781 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     6841 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     6901 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     6961 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     7021 ccagtcga
//
