LOCUS       sequence-456473-        5225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  9 copies of NFAT sites in minimal alphaMHC-pGL-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5225)
  AUTHORS   Wilkins BJ, Dai YS, Bueno OF, Parsons SA, Xu J, Plank DM, Jones F, 
            Kimball TR, Molkentin JD
  TITLE     Calcineurin/NFAT coupling participates in pathological, but not 
            physiological, cardiac hypertrophy.
  JOURNAL   Circ Res. 2004 Jan 9;94(1):110-8. Epub 2003 Dec 1.
  PUBMED    14656927
REFERENCE   2  (bases 1 to 5225)
  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: "Circ Res.";
            date: "2004-01-9"; volume: "94(1)"; pages: "110-8. Epub 2003 Dec 1"
COMMENT     Sequence Label: 9X NFAT-apha-MHC-Luc
FEATURES             Location/Qualifiers
     source          1..5225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      489..498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             495..2147
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(561..579)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2188..2309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2199..2218)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2253..2272
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2557..2574)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2728..3316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2808..2827)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3487..4278,4279..4347))
                     /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     4110..4129
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4348..4452)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4479..4934
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4566..4585)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4776..4797
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5065..5113
                     /note="synthetic polyadenylation signal"
     misc_feature    5127..5218
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5167..5186
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt acattggaaa attttattac actacattgg aaaattttat
       61 tacactacat tggaaaattt tattacacta cattggaaaa ttttattaca cgcgtgctag
      121 cacattggaa aattttatta cacgctagcc ccgcgtacat tggaaaattt tattacacta
      181 cattggaaaa ttttattaca ctacattgga aaattttatt acactacatt ggaaaatttt
      241 attacacgcg gggctgctgt cctcctgtca cctccagagc caagggatca aaggaggagg
      301 agccaggaca ggagggatgg gagggagggt cccagcagat gactccaaat ttaggcagca
      361 ggcacgtgga atgagctata aaggggctgg agcgctgaga gctgtcagac cgagatttct
      421 ccatcccaag cttcggggct cgagatctgc gatctaagta agcttggcat tccggtactg
      481 ttggtaaagc caccatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
      541 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
      601 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
      661 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
      721 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
      781 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
      841 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
      901 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
      961 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
     1021 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
     1081 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
     1141 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
     1201 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
     1261 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
     1321 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
     1381 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
     1441 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
     1501 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg
     1561 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
     1621 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
     1681 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
     1741 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
     1801 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
     1861 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
     1921 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
     1981 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
     2041 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
     2101 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc tagagtcggg
     2161 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2221 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2281 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2341 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2401 aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     2461 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     2521 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2581 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2641 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3781 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4081 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4381 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4441 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     4501 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4561 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4621 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4681 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4741 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4801 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4861 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4921 aacgcttaca atttgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     4981 gcgggcctct tcgctattac gccagcccaa gctaccatga taagtaagta atattaaggt
     5041 acgggaggta cttggagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     5101 ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa
     5161 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat
     5221 cgata
//
