LOCUS Exported 5741 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS NFAT luciferase reporter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5741) AUTHORS Ichida M, Finkel T TITLE Ras regulates NFAT3 activity in cardiac myocytes. JOURNAL J Biol Chem. 2001 Feb 2. 276(5):3524-30. PUBMED 11044444 REFERENCE 2 (bases 1 to 5741) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5741 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(169..186) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(340..928) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(420..439) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1099..1959) /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 1722..1741 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1960..2064) /gene="bla" /label=AmpR promoter rep_origin 2091..2546 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2178..2197) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2388..2409 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 2736..2857 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 2896..2925 /label=NFAT binding site /bound_moiety="NFAT transcription factor" /note="NFAT binding site from the human interleukin 2 (IL-2) gene (Mattila et al., 1990)" protein_bind 2926..2955 /label=NFAT binding site /bound_moiety="NFAT transcription factor" /note="NFAT binding site from the human interleukin 2 (IL-2) gene (Mattila et al., 1990)" protein_bind 2956..2985 /label=NFAT binding site /bound_moiety="NFAT transcription factor" /note="NFAT binding site from the human interleukin 2 (IL-2) gene (Mattila et al., 1990)" CDS 3101..4753 /codon_start=1 /gene="luc" /product="firefly luciferase" /label=luciferase /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL" primer_bind complement(3167..3185) /label=LucNrev /note="Firefly luciferase, reverse primer" intron 4994..5059 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 5189..5209 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 5634..27 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" ORIGIN 1 atcaatgtat cttatcatgt ctggatccgt cgaccgatgc ccttgagagc cttcaaccca 61 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc 121 tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga 181 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 241 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 301 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 361 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 421 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 481 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 541 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 601 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 661 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 721 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 781 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 841 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 901 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 961 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 1021 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 1081 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 1141 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 1201 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 1261 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 1321 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 1381 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 1441 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 1501 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 1561 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 1621 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 1681 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 1741 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 1801 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 1861 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 1921 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 1981 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 2041 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg 2101 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 2161 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 2221 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 2281 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 2341 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 2401 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 2461 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 2521 taacaaaata ttaacgctta caatttgcca ttcgccattc aggctgcgca actgttggga 2581 agggcgatcg gtgcgggcct cttcgctatt acgccagccc aagctaccat gataagtaag 2641 taatattaag gtacgtggag gttttacttg ctttaaaaaa cctcccacac ctccccctga 2701 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg 2761 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 2821 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatgg tactgtaact gagctaacat 2881 aacccgggag gtaccacgcc ttctgtatga aacagttttt cctccacgcc ttctgtatga 2941 aacagttttt cctccacgcc ttctgtatga aacagttttt cctccgggga tctggcgcgg 3001 tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg 3061 cagcgacccg caagcttggc attccggtac tgttggtaaa atggaagacg ccaaaaacat 3121 aaagaaaggc ccggcgccat tctatcctct agaggatgga accgctggag agcaactgca 3181 taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat 3241 cgaggtgaac atcacgtacg cggaatactt cgaaatgtcc gttcggttgg cagaagctat 3301 gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca 3361 attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga 3421 catttataat gaacgtgaat tgctcaacag tatgaacatt tcgcagccta ccgtagtgtt 3481 tgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaaattac caataatcca 3541 gaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt 3601 cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtaccag agtcctttga 3661 tcgtgacaaa acaattgcac tgataatgaa ttcctctgga tctactgggt tacctaaggg 3721 tgtggccctt ccgcatagaa ctgcctgcgt cagattctcg catgccagag atcctatttt 3781 tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt 3841 tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag 3901 atttgaagaa gagctgtttt tacgatccct tcaggattac aaaattcaaa gtgcgttgct 3961 agtaccaacc ctattttcat tcttcgccaa aagcactctg attgacaaat acgatttatc 4021 taatttacac gaaattgctt ctgggggcgc acctctttcg aaagaagtcg gggaagcggt 4081 tgcaaaacgc ttccatcttc cagggatacg acaaggatat gggctcactg agactacatc 4141 agctattctg attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc 4201 attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcagag 4261 aggcgaatta tgtgtcagag gacctatgat tatgtccggt tatgtaaaca atccggaagc 4321 gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag cttactggga 4381 cgaagacgaa cacttcttca tagttgaccg cttgaagtct ttaattaaat acaaaggata 4441 tcaggtggcc cccgctgaat tggaatcgat attgttacaa caccccaaca tcttcgacgc 4501 gggcgtggca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt 4561 ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac 4621 aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac 4681 cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa 4741 gtccaaattg taaaatgtaa ctgtattcag cgatgacgaa attcttagct attgtaatac 4801 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg 4861 cctggttgct acgcctgaat aagtgataat aagcggatga atggcagaaa ttcgccggat 4921 ctttgtgaag gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat 4981 ttaaagctct aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct 5041 aattgtttgt gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa 5101 tgcctttaat gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc 5161 tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa 5221 ggactttcct tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct 5281 tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat 5341 ggaaaaatat tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt 5401 ttttcttact ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg 5461 tacctttagc tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt 5521 gactagagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc 5581 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt 5641 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 5701 catttttttc actgcattct agttgtggtt tgtccaaact c //