LOCUS       sequence-221776-        4852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human dominant-negative 14-3-3 zeta (R56A/R60A) 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4852)
  AUTHORS   Li FQ, Mofunanya A, Harris K, Takemaru K
  TITLE     Chibby cooperates with 14-3-3 to regulate beta-catenin subcellular 
            distribution and signaling activity.
  JOURNAL   J Cell Biol. 2008 Jun 30;181(7):1141-54. doi: 10.1083/jcb.200709091.
            Epub 2008 Jun 23.
  PUBMED    18573912
REFERENCE   2  (bases 1 to 4852)
  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: 
            "10.1083/jcb.200709091"; journalName: "J Cell Biol"; date: 
            "2008-06-30- 30"; volume: "181"; issue: "7"; pages: "1141-54"
COMMENT     Sequence Label: HA-DN-14-3-3 zeta (R56A/R60A)
FEATURES             Location/Qualifiers
     source          1..4852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..999
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1034..1052
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1034..1051
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1091..1117
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1879..1898)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1880..1898)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1903..2037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1927..1946)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1981..2000
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2150..2170)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2150..2168)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2189..2205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2189..2205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2202..2224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2213..2229
                     /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(2237..2243,2244..2261,2262..2267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2282..2303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2420..2437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2591..3179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2671..2690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3350..4141,4142..4210))
                     /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     3973..3992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4211..4315)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4341..4796)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4478..4499)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4690..4709
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca
       61 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
      121 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
      181 atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
      241 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
      301 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
      361 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
      421 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
      481 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
      541 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
      601 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
      661 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
      721 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
      781 gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat
      841 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
      901 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
      961 taatgggagg tctatataag caatgctcgt ttagggaacc gccattctgc ctggggacgt
     1021 cggagcaagc ttgatttagg tgacactata gaatacaagc tacttgttct ttttgcagga
     1081 taccatgcga tacccatatg atgtgccaga ttacgctgaa ttcaccatgg ataaaaatga
     1141 gctggttcag aaggccaaac tggccgagca ggctgagcga tatgatgaca tggcagcctg
     1201 catgaagtct gtaactgagc aaggagctga attatccaat gaggagagga atcttctctc
     1261 agttgcttat aaaaatgttg taggagcccg tgcgtcatct tgggcggtcg tctcaagtat
     1321 tgaacaaaag acggaaggtg ctgagaaaaa acagcagatg gctcgagaat acagagagaa
     1381 aattgagacg gagctaagag atatctgcaa tgatgtactg tctcttttgg aaaagttctt
     1441 gatccccaat gcttcacaag cagagagcaa agtcttctat ttgaaaatga aaggagatta
     1501 ctaccgttac ttggctgagg ttgccgctgg tgatgacaag aaagggattg tcgatcagtc
     1561 acaacaagca taccaagaag cttttgaaat cagcaaaaag gaaatgcaac caacacatcc
     1621 tatcagactg ggtctggccc ttaacttctc tgtgttctat tatgagattc tgaactcccc
     1681 agagaaagcc tgctctcttg caaagacagc ttttgatgaa gccattgctg aacttgatac
     1741 attaagtgaa gagtcataca aagacagcac gctaataatg caattactga gagacaactt
     1801 gacattgtgg acatcggata cccaaggaga cgaagctgaa gcaggagaag gaggggaaaa
     1861 ttaatctaga ttctgcagcc ctatagtgag tcgtattacg tagatccaga catgataaga
     1921 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1981 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2041 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     2101 ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
     2161 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2221 cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct
     2281 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2341 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2401 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2461 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2521 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2581 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2641 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2701 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2761 cttcgggaaa cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2821 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2881 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2941 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3001 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3061 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3121 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3181 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3241 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3301 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3361 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3421 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3481 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3541 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3601 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3661 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3721 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3781 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3841 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3901 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3961 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4021 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4081 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4141 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4201 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4261 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4321 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4381 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4441 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4501 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4561 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4621 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4681 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4741 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     4801 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct ct
//
