LOCUS       sequence-221775-        4852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human 14-3-3 zeta 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-14-3-3 zeta
FEATURES             Location/Qualifiers
     source          1..4852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        86..1070
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1105..1123
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1105..1122
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1162..1188
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1950..1969)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1951..1969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1974..2108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1998..2017)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2052..2071
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2221..2241)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2221..2239)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2260..2276)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2260..2276)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2273..2295)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2284..2300
                     /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(2308..2314,2315..2332,2333..2338))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2353..2374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2491..2508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2662..3250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2742..2761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3421..4212,4213..4281))
                     /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     4044..4063
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4282..4386)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(4412..4852,1..15))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4549..4570)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4761..4780
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
       61 tgcgggcctc ttcgctatta cgccagtcga ccgccaattc aatatggcgt atatggactc
      121 atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc
      181 gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc
      241 atgccaattc aatatggcgg acctggcact gtgccaactg gggaggggtc tacttggcac
      301 ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat
      361 ggtgccaata atggcggcca tattggctat atgccaggat caatatatag gcaatatcca
      421 atatggccct atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc
      481 catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg
      541 tcccatatac catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc
      601 aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata
      661 tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg
      721 acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc
      781 gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat
      841 atctattaat agtaacttgg caagtacatt actattggaa gtacgccagg gtacattggc
      901 agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc
      961 attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg
     1021 gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac cgccattctg
     1081 cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag ctacttgttc
     1141 tttttgcagg ataccatgcg atacccatat gatgtgccag attacgctga attcaccatg
     1201 gataaaaatg agctggttca gaaggccaaa ctggccgagc aggctgagcg atatgatgac
     1261 atggcagcct gcatgaagtc tgtaactgag caaggagctg aattatccaa tgaggagagg
     1321 aatcttctct cagttgctta taaaaatgtt gtaggagccc gtaggtcatc ttggagggtc
     1381 gtctcaagta ttgaacaaaa gacggaaggt gctgagaaaa aacagcagat ggctcgagaa
     1441 tacagagaga aaattgagac ggagctaaga gatatctgca atgatgtact gtctcttttg
     1501 gaaaagttct tgatccccaa tgcttcacaa gcagagagca aagtcttcta tttgaaaatg
     1561 aaaggagatt actaccgtta cttggctgag gttgccgctg gtgatgacaa gaaagggatt
     1621 gtcgatcagt cacaacaagc ataccaagaa gcttttgaaa tcagcaaaaa ggaaatgcaa
     1681 ccaacacatc ctatcagact gggtctggcc cttaacttct ctgtgttcta ttatgagatt
     1741 ctgaactccc cagagaaagc ctgctctctt gcaaagacag cttttgatga agccattgct
     1801 gaacttgata cattaagtga agagtcatac aaagacagca cgctaataat gcaattactg
     1861 agagacaact tgacattgtg gacatcggat acccaaggag acgaagctga agcaggagaa
     1921 ggaggggaaa attaatctag attctgcagc cctatagtga gtcgtattac gtagatccag
     1981 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2041 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2101 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2161 aggtttttta attcgcggcc gcggcgccaa tgcattgggc ccggtaccca gcttttgttc
     2221 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2281 aaattgttat ccgctcacaa ttccacacaa catacgagcc gggagcataa agtgtaaagc
     2341 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2401 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2461 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2521 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2581 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2641 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2701 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2761 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2821 cgcctttctc ccttcgggaa acgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2881 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2941 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3001 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3061 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3121 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3181 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3241 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3301 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3361 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3421 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3481 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3541 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3601 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3661 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3721 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3781 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3841 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3901 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3961 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4021 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4081 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4141 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4201 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4261 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4321 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4381 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     4441 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4501 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4561 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4621 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4681 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4741 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4801 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gc
//
