LOCUS Exported 5123 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PUFb_p65HSF1 in pmax-DEST expression vector. ACCESSION . VERSION . KEYWORDS pAC1392_pmax-PUFb_p65HSF1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5123) AUTHORS Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, Taghbalout A, Wang H TITLE Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. JOURNAL Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15. PUBMED 26768771 REFERENCE 2 (bases 1 to 5123) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5123 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 138..517 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 518..721 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 672..692 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" intron 813..945 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" protein_bind 960..984 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 1002..1011 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1023..1043 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2154..2174 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2190..2732 /codon_start=1 /product="C-terminal portion of the p65 subunit of mouse NF-kappa-B" /label=p65 /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED FSSIADMDFSALLSQISS" CDS 2757..3128 /codon_start=1 /product="C-terminal activation domain from the human heat shock transcription factor HSF1" /label=HSF1 /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED PTISLLTGSEPPKAKDPTVS" protein_bind complement(3153..3177) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3230..3271 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3297..3316) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" polyA_signal 3326..3407 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 3351..3370 /label=SV40pA-R /note="SV40 polyA, reverse primer" rep_origin complement(3605..4192) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3685..3704) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4273..5067) /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4384..4403) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4994..5013 /label=Neo-R /note="Neomycin resistance gene, reverse 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 gtaataaccc 661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg 721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat 781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt 841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg 901 cacctattgg tcttactgac atccactttg cctttctctc cacagggaaa aaacaattga 961 caagtttgta caaaaaagca ggctccgaat tcaccggtgc cgccaccatg gggatcctcc 1021 cccccaagaa aaagaggaag gtatctagag gccgcagccg ccttttggaa gattttcgaa 1081 acaaccggta ccccaattta caactgcggg agattgctgg acatataatg gaattttccc 1141 aagaccagca tgggtccaga ttcattcagc tgaaactgga gcgtgccaca ccagctgagc 1201 gccagcttgt cttcaatgaa atcctccagg ctgcctacca actcatggtg gatgtgtttg 1261 gtaattacgt cattcagaag ttctttgaat ttggcagtct tgaacagaag ctggctttgg 1321 cagaacggat tcgaggccac gtcctgtcat tggcactaca gatgtatggc tgccgtgtta 1381 tccagaaagc tcttgagttt attccttcag accagcagaa tgagatggtt cgggaactag 1441 atggccatgt cttgaagtgt gtgaaagatc agaatggcaa tcacgtggtt cagaaatgca 1501 ttgaatgtgt acagccccag tctttgcaat ttatcatcga tgcgtttaag ggacaggtat 1561 ttgccttatc cacacatcct tatggctgcc gagtgattca gagaatcctg gagcactgtc 1621 tccctgacca gacactccct attttagagg agcttcacca gcacacagag cagctggtac 1681 aggatcaata tggaagttat gtaatcgaac atgtactgga gcacggtcgt cctgaggata 1741 aaagcaaaat tgtagcagaa atccgaggca atgtacttgt attgagtcag cacaaatttg 1801 caaacaatgt tgtgcagaag tgtgttactc acgcctcacg tacggagcgc gctgtgctca 1861 tcgatgaggt gtgcaccatg aacgacggtc cccacagtgc cttatacacc atgatgaagg 1921 accagtatgc caactacgtg gtccagaaga tgattgacgt ggcggagcca ggccagcgga 1981 agatcgtcat gcataagatc cggccccaca tcgcaactct tcgtaagtac acctatggca 2041 agcacattct ggccaagctg gagaagtact acatgaagaa cggtgttgac ttaggggggc 2101 cggccggagg aggtggaagc ggaggaggag gaagcggagg aggaggtagc ggacctaaga 2161 aaaagaggaa ggtggcggcc gctggatccc cttcagggca gatcagcaac caggccctgg 2221 ctctggcccc tagctccgct ccagtgctgg cccagactat ggtgccctct agtgctatgg 2281 tgcctctggc ccagccacct gctccagccc ctgtgctgac cccaggacca ccccagtcac 2341 tgagcgctcc agtgcccaag tctacacagg ccggcgaggg gactctgagt gaagctctgc 2401 tgcacctgca gttcgacgct gatgaggacc tgggagctct gctggggaac agcaccgatc 2461 ccggagtgtt cacagatctg gcctccgtgg acaactctga gtttcagcag ctgctgaatc 2521 agggcgtgtc catgtctcat agtacagccg aaccaatgct gatggagtac cccgaagcca 2581 ttacccggct ggtgaccggc agccagcggc cccccgaccc cgctccaact cccctgggaa 2641 ccagcggcct gcctaatggg ctgtccggag atgaagactt ctcaagcatc gctgatatgg 2701 actttagtgc cctgctgtca cagatttcct ctagtgggca gggaggaggt ggaagcggct 2761 tcagcgtgga caccagtgcc ctgctggacc tgttcagccc ctcggtgacc gtgcccgaca 2821 tgagcctgcc tgaccttgac agcagcctgg ccagtatcca agagctcctg tctccccagg 2881 agccccccag gcctcccgag gcagagaaca gcagcccgga ttcagggaag cagctggtgc 2941 actacacagc gcagccgctg ttcctgctgg accccggctc cgtggacacc gggagcaacg 3001 acctgccggt gctgtttgag ctgggagagg gctcctactt ctccgaaggg gacggcttcg 3061 ccgaggaccc caccatctcc ctgctgacag gctcggagcc tcccaaagcc aaggacccca 3121 ctgtctccat cgattgatta attaagaatt cgacccagct ttcttgtaca aagtggttga 3181 tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat 3241 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt tagcaattgt tttttcgatg 3301 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg 3361 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt 3421 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa 3481 acctctacaa atgtggtact taagaggggg agaccaaagg gcgagacgtt aaggcctcac 3541 gtgacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 3601 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 3661 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 3721 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 3781 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 3841 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 3901 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 3961 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 4021 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 4081 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacg ctggtagcgg 4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 4261 ggtcatgccg tctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc 4321 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc 4381 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc 4441 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc 4501 gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag 4561 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc 4621 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt 4681 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc 4741 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc 4801 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag 4861 ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt 4921 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc 4981 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc 5041 tgcgtgcaat ccatcttgtt caatcataat attattgaag catttatcag ggttcgtctc 5101 gtcccggtct cctcccatgc atg //