LOCUS       sequence-194471-        7762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human HA-14-3-3 ϵ (epsilon) and myc-14-3-3-zeta (zeta) in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7762)
  AUTHORS   Chen HK, Fernandez-Funez P, Acevedo SF, Lam YC, Kaytor MD, Fernandez
            MH, Aitken A, Skoulakis EM, Orr HT, Botas J, Zoghbi HY
  TITLE     Interaction of Akt-phosphorylated ataxin-1 with 14-3-3 mediates 
            neurodegeneration in spinocerebellar ataxia type 1.
  JOURNAL   Cell. 2003 May 16;113(4):457-68.
  PUBMED    12757707
REFERENCE   2  (bases 1 to 7762)
  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: "Cell."; 
            date: "2003-05-16"; volume: "113(4)"; pages: "457-68"
COMMENT     Sequence Label: pIRES-HA-14-3-3 ϵ/myc-14-3-3-zeta
FEATURES             Location/Qualifiers
     source          1..7762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1105..1131)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1949..1975
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(2035..2609,2610..2612,2613..2621)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2202..2219)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2429..2448
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3371..3400
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3466..3587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3477..3496)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3531..3550
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3773..4228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3860..3879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4070..4091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4345..4702
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4345..4536
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4368..4388)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4553..4688
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4753..5547
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4807..4826)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5417..5436
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5611..5659
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5700..5719)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5819..5841
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5879..5897)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5965..6069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6070..6138,6139..6930)
                     /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     complement(6288..6307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7101..7689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7590..7609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagaatt ctcaagcata atctggaaca tcatatggat actgattttc
     1141 gtcttccacc cgtcgcgcag accctgccgg agccgctgcc gctatggatg atcgagagga
     1201 tctggtgtac caggcgaagc tggccgagca ggctgagcga tacgacgaaa tggtggagtc
     1261 aatgaagaaa gtagcaggga tggatgtgga gctgacagtt gaagaaagaa acctcctatc
     1321 tgttgcatat aagaatgtga ttggagctag aagagcctcc tggagaataa tcagcagcat
     1381 tgaacagaaa gaagaaaaca agggaggaga agacaagcta aaaatgattc gggaatatcg
     1441 gcaaatggtt gagactgagc taaagttaat ctgttgtgac attctggatg tactggacaa
     1501 acacctcatt ccagcagcta acactggcga gtccaaggtt ttctattata aaatgaaagg
     1561 ggactaccac aggtatctgg cagaatttgc cacaggaaac gacaggaagg aggctgcgga
     1621 gaacagccta gtggcttata aagctgctag tgatattgca atgacagaac ttccaccaac
     1681 gcatcctatt cgcttaggtc ttgctctcaa tttttccgta ttctactacg aaattcttaa
     1741 ttcccctgac cgtgcctgca ggttggcaaa agcagctttt gatgatgcaa ttgcagaact
     1801 ggatacgctg agtgaagaaa gctataagga ctctacactt atcatgcagt tgttacgtga
     1861 taatctgaca ctatggactt cagacatgca gggtgacggt gaagagcaga ataaagaagc
     1921 gctgcaggac gtggaagacg aaaatcagta tccatatgat gttccagatt atgcttgaga
     1981 attcacgcgt cgagcatgca tctagggcgg ccaattccgc ccctctcccc cccccccctc
     2041 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     2101 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     2161 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     2221 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     2281 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     2341 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     2401 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     2461 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     2521 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     2581 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgggatcct ctagaatgga
     2641 taaaaatgag ctggttcaga aggccaaact ggccgagcag gctgagcgat atgatgacat
     2701 ggcagcctgc atgaagtctg taactgagca aggagctgaa ttatccaatg aggagaggaa
     2761 tcttctctca gttgcttata aaaatgttgt aggagcccgt aggtcatctt ggagggtcgt
     2821 ctcaagtatt gaacaaaaga cggaaggtgc tgagaaaaaa cagcagatgg ctcgagaata
     2881 cagagagaaa attgagacgg agctaagaga tatctgcaat gatgtactgt ctcttttgga
     2941 aaagttcttg atccccaatg cttcacaagc agagagcaaa gtcttctatt tgaaaatgaa
     3001 aggagattac taccgttact tggctgaggt tgccgctggt gatgacaaga aagggattgt
     3061 cgatcagtca caacaagcat accaagaagc ttttgaaatc agcaaaaagg aaatgcaacc
     3121 aacacatcct atcagactgg gtctggccct taacttctct gtgttctatt atgagattct
     3181 gaactcccca gagaaagcct gctctcttgc aaagacagct tttgatgaag ccattgctga
     3241 acttgataca ttaagtgaag agtcatacaa agacagcacg ctaataatgc aattactgag
     3301 agacaacttg acattgtgga catcggatac ccaaggagac gaagctgaag caggagaagg
     3361 aggggaaaat gaacaaaaac tcatctcaga agaggatctg taatctagag tcgacccggg
     3421 cggccgcttc cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg
     3481 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3541 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3601 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     3661 acctctacaa atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga
     3721 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     3781 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3841 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3901 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3961 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4021 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4081 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4141 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4201 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     4261 gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga
     4321 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     4381 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4441 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4501 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4561 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4621 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4681 tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag
     4741 gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4801 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4861 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4921 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4981 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5041 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5101 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5161 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5221 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5281 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5341 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5401 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5461 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5521 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5581 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     5641 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt
     5701 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     5761 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5821 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     5881 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5941 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6001 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6061 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6121 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6181 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6241 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6301 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6361 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6421 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6481 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6541 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6601 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6661 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6721 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6781 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6841 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6901 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6961 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7021 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7081 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7741 gctcacatgg ctcgacagat ct
//
