LOCUS       sequence-221771-        4863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human 14-3-3 eta in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4863)
  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 4863)
  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-14-3-3 eta
FEATURES             Location/Qualifiers
     source          1..4863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..1013
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1048..1066
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1048..1065
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1105..1131
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1904..1923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1905..1923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1928..2062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1952..1971)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2006..2025
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2175..2195)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2175..2193)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2214..2230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2214..2230)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2227..2249)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2238..2254
                     /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(2262..2268,2269..2286,2287..2292))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2307..2328
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2445..2462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2616..3204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2696..2715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3375..4166,4167..4235))
                     /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     3998..4017
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4236..4340)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4366..4821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4503..4524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4715..4734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga
       61 ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga
      121 ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc
      181 accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg
      241 cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg
      301 catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat
      361 ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta
      421 tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg
      481 cggtcccata taccatatat ggggcttcct aataccgccc atagccactc ccccattgac
      541 gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt
      601 atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca
      661 ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg cccattcata
      721 tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc
      781 aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt
      841 ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc
      901 cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg
      961 gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aaccgccatt
     1021 ctgcctgggg acgtcggagc aagcttgatt taggtgacac tatagaatac aagctacttg
     1081 ttctttttgc aggataccat gcgataccca tatgatgtgc cagattacgc tgaattcacc
     1141 atgggggacc gggagcagct gctgcagcgg gcgcggctgg ccgagcaggc ggagcgctac
     1201 gacgacatgg cctccgctat gaaggcggtg acagagctga atgaacctct ctccaatgaa
     1261 gatcgaaatc tcctctctgt ggcctacaag aatgtggttg gtgccaggcg atcttcctgg
     1321 agggtcatta gcagcattga gcagaaaacc atggctgatg gaaacgaaaa gaaattggag
     1381 aaagttaaag cttaccggga gaagattgag aaggagctgg agacagtttg caatgatgtc
     1441 ctgtctctgc ttgacaagtt cctgatcaag aactgcaatg atttccagta tgagagcaag
     1501 gtgttttacc tgaaaatgaa gggtgattac taccgctact tagcagaggt cgcttctggg
     1561 gagaagaaaa acagtgtggt cgaagcttct gaagctgcct acaaggaagc ctttgaaatc
     1621 agcaaagagc agatgcaacc cacgcatccc atccggctgg gcctggccct caacttctcc
     1681 gtgttctact atgagatcca gaatgcacct gagcaagcct gcctcttagc caaacaagcc
     1741 ttcgatgatg ccatagctga gctggacaca ctaaacgagg attcctataa ggactccacg
     1801 ctgatcatgc agttgctgcg agacaacctc accctctgga cgagcgacca gcaggatgaa
     1861 gaagcaggag aaggcaactg actcgagcct ctagattctg cagccctata gtgagtcgta
     1921 ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1981 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2041 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2101 gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta
     2161 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     2221 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc
     2281 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2341 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2401 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2461 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2521 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2761 accggatacc tgtccgcctt tctcccttcg ggaaacgtgg cgctttctca tagctcacgc
     2821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3301 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3361 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3421 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3481 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3541 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3601 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3661 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3721 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3781 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3841 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3901 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3961 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4021 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4081 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4141 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4201 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4261 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4321 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     4381 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4441 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4501 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4561 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4621 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4681 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4741 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4801 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     4861 gat
//
