LOCUS Exported 5157 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Four BHLHE40 consensus binding sites cloned upstream of luciferase reporter gene. ACCESSION . VERSION . KEYWORDS pGL3-Promoter-4xBS_BHLHE40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5157) AUTHORS Emming S, Bianchi N, Polletti S, Balestrieri C, Leoni C, Montagner S, Chirichella M, Delaleu N, Natoli G, Monticelli S TITLE A molecular network regulating the proinflammatory phenotype of human memory T lymphocytes. JOURNAL Nat Immunol. 2020 Apr;21(4):388-399. doi: 10.1038/s41590-020-0622-8. Epub 2020 Mar 16. PUBMED 32205878 REFERENCE 2 (bases 1 to 5157) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5157 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 195..391 /label=SV40 promoter /note="SV40 early promoter" rep_origin 242..377 /label=SV40 ori /note="SV40 origin of replication" primer_bind 304..323 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" regulatory 421..430 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 427..2079 /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(493..511) /label=LucNrev /note="Firefly luciferase, reverse primer" polyA_signal 2120..2241 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2131..2150) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2185..2204 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2489..2506) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2660..3248) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2740..2759) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3419..4279) /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 4042..4061 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4280..4384) /gene="bla" /label=AmpR promoter rep_origin 4411..4866 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4498..4517) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4708..4729 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 4997..5045 /note="synthetic polyadenylation signal" misc_feature 5059..5150 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 5099..5118 /label=RVprimer3 /note="pGL3 vector, forward primer" ORIGIN 1 ggtaccgagc gattacgcgt gctagcaatc cttgtgtcac gtgacacaag gattgctaga 61 cgtcacgtga cacaaggatt ctagcccggg atcgagctct tacgcgtgct agcaatcctt 121 gtgtcacgtg acacaaggat tgctagacgt cacgtgacac aaggattgct agcccgggct 181 cgagatctgc gatctgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 241 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatcgct gactaatttt 301 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 361 aggctttttt ggaggcctag gcttttgcaa aaagcttggc attccggtac tgttggtaaa 421 gccaccatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta tccgctggaa 481 gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct ggttcctgga 541 acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga gtacttcgaa 601 atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa tcacagaatc 661 gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc gttatttatc 721 ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct caacagtatg 781 ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac 841 gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa aacggattac 901 cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg ttttaatgaa 961 tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat catgaactcc 1021 tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga 1081 ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac tgcgatttta 1141 agtgttgttc cattccatca cggttttgga atgtttacta cactcggata tttgatatgt 1201 ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag gagccttcag 1261 gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt cgccaaaagc 1321 actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg tggcgctccc 1381 ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg tatcaggcaa 1441 ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg ggatgataaa 1501 ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga tctggatacc 1561 gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg 1621 tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga tggatggcta 1681 cattctggag acatagctta ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg 1741 aagtctctga ttaagtacaa aggctatcag gtggctcccg ctgaattgga atccatcttg 1801 ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt 1861 gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga aaaagagatc 1921 gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt 1981 gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat cagagagatc 2041 ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat tctagagtcg gggcggccgg 2101 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg 2161 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt 2221 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag 2281 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat 2341 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc 2401 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg 2461 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 2521 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 2581 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 2641 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 2701 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 2761 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 2821 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 2881 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 2941 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 3001 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 3061 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 3121 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 3181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 3241 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 3301 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 3361 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 3421 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 3481 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 3541 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 3601 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 3661 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 3721 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 3781 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 3841 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 3901 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 3961 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 4021 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 4081 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 4141 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 4201 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 4261 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 4321 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 4381 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg 4441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 4501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 4561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 4621 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 4681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 4741 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 4801 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta 4861 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct 4921 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg 4981 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt 5041 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact 5101 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgata //