LOCUS Exported 9423 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS WT NFAT1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9423) AUTHORS Monticelli S, Rao A TITLE NFAT1 and NFAT2 are positive regulators of IL-4 gene transcription. JOURNAL Eur J Immunol. 2002 Oct . 32(10):2971-8. PUBMED 12355451 REFERENCE 2 (bases 1 to 9423) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9423 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /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 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter primer_bind 1974..1992 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2030..2052) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2152..2171 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2365..2387 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2515..2534 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 2734..3250 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 3314..3655 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 3722..4138 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 4068..4090 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 4106..4122 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" protein_bind 4163..4187 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 4232..4321 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA" protein_bind complement(7148..7172) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 7220..7772 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(7364..7381) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 7591..7610 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 7784..8503 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(7829..7850) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(8090..8109) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 8437..8458 /label=EGFP-C /note="EGFP, forward primer" LTR 8550..9065 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 9227..9243 /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(9251..9281) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9296..9317 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 1981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 2041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 2161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 2221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 2281 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 2341 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa 2401 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc 2461 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat 2521 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg 2581 tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct 2641 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata 2701 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg 2761 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag 2821 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg 2881 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc 2941 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat 3001 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt 3061 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga 3121 tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa 3181 tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg 3241 gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc 3301 cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt 3361 ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat 3421 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg 3481 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa 3541 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc 3601 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg 3661 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta 3721 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg 3781 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc 3841 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat 3901 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc 3961 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt 4021 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc 4081 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg 4141 aattagatct ctcgaggtca tcacaagttt gtacaaaaaa gcaggcttcg aaggagatag 4201 aaccaattct taagcttcca ccatgatctt ttacccatac gatgttcctg actatgcggg 4261 ctatccctat gacgtcccgg actatgcagg atcctatcca tatgacgttc cagattacgc 4321 tgctcagtgc ggccgcgacg tcccggagcc gcagcccgac cccgatggcg gggacggccc 4381 cggccacgag cccgggggca gtccccaaga cgagctggac ttttccatcc tcttcgatta 4441 tgactatctg aaccctatcg aagaagaacc gatcgcacat aaggccatca gctcaccctc 4501 cggactcgca tacccggatg atgtcctgga ctatggcctc aagccatgca acccccttgc 4561 cagtctctct ggcgagcccc ctggccggtt cggagagccg gatagtatag ggttccagaa 4621 ctttctgagc ccggtcaagc cagcaggggc ttcgggcccg agccctcgga tcgagatcac 4681 tccatcccac gaactgatgc aggcaggggg ggccctccgt gggagagacg ccggcctgtc 4741 ccccgagcag ccggccctgg ccctggccgg cgtggccgcc agcccgaggt tcacactgcc 4801 cgtgcccggc tacgagggct accgcgagcc gctttgcttg agccccgcta gcagcggctc 4861 ctctgccagc ttcatttctg acaccttctc cccctacacc tcgccctgcg tctcacccaa 4921 taacgccggg cccgacgacc tgtgtcccca gtttcaaaac atccctgctc attattcccc 4981 cagaacctct ccaataatgt cacctcgaac cagcctcgcc gaggacagct gcctgggccg 5041 acactcgccc gtgccccgtc cggcatcccg ctcctcctca cccggtgcca agcggaggca 5101 ttcgtgcgca gaggctttgg ttgctcctct gcccgcagcc tcaccccagc gctcccggag 5161 cccctcgcca cagccctcgc ctcacgtggc accgcaggac gacagcatcc ccgctgggta 5221 cccccccacg gccggctctg ctgttctcat ggatgccctc aacaccctgg ccaccgactc 5281 gccctgcggg atcccctcca agatatggaa gaccagtcct gacccgacgc ctgtgtccac 5341 cgctccgtcc aaggctggcc tggcccgcca catctaccct actgtggagt tcctggggcc 5401 atgtgagcag gaggagagga ggaattccgc tccagagtcc atcctgctgg taccacctac 5461 ttggcccaag cagttggtgc cggccattcc catctgcagc atccctgtga ctgcatccct 5521 cccaccactc gagtggccac tctccaatca gtcgggctcc tatgagctac ggattgaggt 5581 ccaacccaag ccccatcacc gggcccacta tgagacggag ggcagccgtg gcgctgtcaa 5641 agccccaaca ggaggacacc ctgtggtgca gctccacggc tacatggaga acaagcctct 5701 ggggcttcag atcttcattg ggacagcaga tgagaggatc cttaagccgc acgccttcta 5761 ccaagtacac aggatcactg ggaaaacggt caccaccacg agctatgaga agatcgtagg 5821 caacaccaag gtcctggaga tccccctgga gccaaagaac aacatgagag ccaccatcga 5881 ctgtgcaggc atcctgaagc tccgaaacgc tgacatcgag ctgcggaagg gcgagacgga 5941 catcggcagg aagaacacgc gtgtgcgcct ggtgttccgc gtgcacgtcc cagagcccag 6001 tgggcgcatc gtctccctgc aggctgcgtc caaccccatc gagtgctctc agcgctctgc 6061 ccacgagctg cccatggtgg agagacaaga catggacagc tgcctggtct acgggggcca 6121 gcagatgatc ctcacgggcc agaacttcac agcggagtcc aaggttgtgt tcatggagaa 6181 gactacagat gggcagcaga tttgggagat ggaagctacg gtggataaag acaagagcca 6241 gcctaacatg ctttttgttg agatccccga gtatcggaac aagcacatcc gcgtgcccgt 6301 gaaagtcaac ttctacgtca tcaacggaaa gaggaaacga agtcagccac agcactttac 6361 ctaccaccca gtccctgcca tcaagacaga gcccagcgat gagtatgaac catctttgat 6421 ctgcagcccc gcccatggag gcctggggag ccagccatat tacccacagc acccaatgct 6481 ggccgagtcc ccctcctgcc ttgtggctac catggccccc tgccaacagt tccgctcggg 6541 gctctcatcc cccgatgctc gctaccaaca gcagagcccc gcagctgccc tctaccagag 6601 aagcaagagc ctgagtcccg gcctgctggg ctaccagcag ccgtccctcc tggcagcacc 6661 cttgggtctg gctgatgccc accgctctgt gctggtgcat gctggttctc aggggcaggg 6721 gcagggctcc accctccgac acacatcctc ggccagccag caggcctcac ccgtgatcca 6781 ctactcaccc accaaccagc agcttcgcgg tgggggtcac caggagttcc agcatatcat 6841 gtactgtgaa aacttcggcc ccagctctgc caggcctggc ccgcctccca tcaaccaagg 6901 tcagaggctg agcccgggcg cctaccccac agtcatccaa caacagactg ccccgagcca 6961 aagagctgcc aaaaacggac ccagtgacca gaaggaagct ctgcccacgg gagtgaccgt 7021 caaacaggaa cagaacctgg accagaccta cctggatgac gttaatgaaa tcatcaggaa 7081 ggagttttca ggacctccct cccgaaatca gacctagaaa caaacataat ctcgcgacgc 7141 aatctagacc cagctttctt gtacaaagtg gtgatgacgg tatcgataag cttgatatcg 7201 aattccgccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg 7261 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa 7321 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg 7381 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca 7441 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc 7501 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt 7561 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg 7621 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca 7681 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac 7741 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga 7801 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca 7861 caagttcagc gtgtctggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa 7921 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac 7981 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa 8041 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa 8101 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct 8161 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta 8221 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggcgaactt 8281 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa 8341 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc 8401 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac 8461 cgccgccggg atcactctcg gcatggacga gctgtacaag taatgagata aaataaaaga 8521 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc 8581 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt 8641 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta 8701 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct 8761 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc 8821 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct 8881 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac 8941 tgcgtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgacttgtg 9001 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct 9061 ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg agtcgaatat 9121 taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat actcctgaaa 9181 tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt tatccgctca 9241 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 9301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 9361 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 9421 gct //