LOCUS       sequence-221769-        4890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human 14-3-3 epsilon in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4890)
  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 4890)
  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 epsilon
FEATURES             Location/Qualifiers
     source          1..4890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        60..1044
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1079..1097
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1079..1096
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1136..1162
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1962..1981)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1963..1981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1986..2120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2010..2029)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2064..2083
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2233..2253)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2233..2251)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2272..2288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2272..2288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2285..2307)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2296..2312
                     /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(2320..2326,2327..2344,2345..2350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2365..2386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2503..2520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2674..3262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2754..2773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3433..4224,4225..4293))
                     /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     4056..4075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4294..4398)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4424..4879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4561..4582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4773..4792
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
       61 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
      121 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
      181 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg
      241 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
      301 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
      361 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
      421 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
      481 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
      541 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
      601 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
      661 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
      721 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
      781 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
      841 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
      901 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
      961 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
     1021 ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag caagcttgat
     1081 ttaggtgaca ctatagaata caagctactt gttctttttg caggatacca tgcgataccc
     1141 atatgatgtg ccagattacg ctgaattcac catggatgat cgagaggatc tggtgtacca
     1201 ggcgaagctg gccgagcagg ctgagcgata cgacgaaatg gtggagtcaa tgaagaaagt
     1261 agcagggatg gatgtggagc tgacagttga agaaagaaac ctcctatctg ttgcatataa
     1321 gaatgtgatt ggagctagaa gagcctcctg gagaataatc agcagcattg aacagaaaga
     1381 agaaaacaag ggaggagaag acaagctaaa aatgattcgg gaatatcggc aaatggttga
     1441 gactgagcta aagttaatct gttgtgacat tctggatgta ctggacaaac acctcattcc
     1501 agcagctaac actggcgagt ccaaggtttt ctattataaa atgaaagggg actaccacag
     1561 gtatctggca gaatttgcca caggaaacga caggaaggag gctgcggaga acagcctagt
     1621 ggcttataaa gctgctagtg atattgcaat gacagaactt ccaccaacgc atcctattcg
     1681 cttaggtctt gctctcaatt tttccgtatt ctactacgaa attcttaatt cccctgaccg
     1741 tgcctgcagg ttggcaaaag cagcttttga tgatgcaatt gcagaactgg atacgctgag
     1801 tgaagaaagc tataaggact ctacacttat catgcagttg ttacgtgata atctgacact
     1861 atggacttca gacatgcagg gtgacggtga agagcagaat aaagaagcgc tgcaggacgt
     1921 ggaagacgaa aatcagtgac tcgagcctct agattctgca gccctatagt gagtcgtatt
     1981 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2041 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2101 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2161 gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc
     2221 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     2281 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat
     2341 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2401 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2461 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2521 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2581 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2641 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2701 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2761 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2821 cggatacctg tccgcctttc tcccttcggg aaacgtggcg ctttctcata gctcacgctg
     2881 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2941 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3001 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3061 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3121 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3181 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3241 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3661 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3721 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3781 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3841 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3901 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3961 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4021 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4081 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4141 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4201 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4261 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     4441 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4501 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4561 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4621 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4681 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4741 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4801 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4861 gcgccgctac agggcgcgtc ccattcgcca
//
