LOCUS Exported 6656 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION 4xNLS_PUFa_2xNLS_mCBPHAT in pmax expression vector. ACCESSION . VERSION . KEYWORDS pAC1417-pmax-4xNLS_PUFa_2xNLS_mCBPHAT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6656) 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 6656) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6656 /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" CDS 1035..1055 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1059..1079 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1083..1103 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1194..1214 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2271..2291 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2295..2315 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" regulatory 3996..4005 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(4686..4710) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 4763..4804 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(4830..4849) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" polyA_signal 4859..4940 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 4884..4903 /label=SV40pA-R /note="SV40 polyA, reverse primer" rep_origin complement(5138..5725) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5218..5237) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5806..6600) /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(5917..5936) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 6527..6546 /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 atcgatggcg 1021 gaggcggatc tgatccaaaa aagaagagaa aggtagatcc aaaaaagaag agaaaggtag 1081 atccaaaaaa gaagagaaag gtaggatcta ccggatctag aaacgatggt ggtggtggaa 1141 gcgggggtgg gggcagcggt ggagggggaa gcgggcgcgc cgggatcctc ccccccaaga 1201 aaaagaggaa ggtatctaga ggccgcagcc gccttttgga agattttcga aacaaccggt 1261 accccaattt acaactgcgg gagattgctg gacatataat ggaattttcc caagaccagc 1321 atgggtccag attcattcag ctgaaactgg agcgtgccac accagctgag cgccagcttg 1381 tcttcaatga aatcctccag gctgcctacc aactcatggt ggatgtgttt ggtaattacg 1441 tcattcagaa gttctttgaa tttggcagtc ttgaacagaa gctggctttg gcagaacgga 1501 ttcgaggcca cgtcctgtca ttggcactac agatgtatgg cagccgtgtt atcgagaaag 1561 ctcttgagtt tattccttca gaccagcaga atgagatggt tcgggaacta gatggccatg 1621 tcttgaagtg tgtgaaagat cagaatggca atcacgtggt tcagaaatgc attgaatgtg 1681 tacagcccca gtctttgcaa tttatcatcg acgcgtttaa gggacaggta tttgccttat 1741 ccacacatcc ttatggctgc cgagtgattc agagaatcct ggagcactgt ctccctgacc 1801 agacactccc tattttagag gagcttcacc agcacacaga gcagctggta caggatcaat 1861 atggaaatta tgtaatccaa catgtactgg agcacggtcg tcctgaggat aaaagcaaaa 1921 ttgtagcaga aatccgaggc aatgtacttg tattgagtca gcacaaattt gcaagcaatg 1981 ttgtggagaa gtgtgttact cacgcctcac gtacggagcg cgctgtgctc atcgacgagg 2041 tgtgcaccat gaacgacggt ccccacagtg ccttatacac catgatgaag gaccagtatg 2101 ccaactacgt ggtccagaag atgattgacg tggcggagcc aggccagcgg aagatcgtca 2161 tgcataagat ccggccccac atcgcaactc ttcgtaagta cacctatggc aagcacattc 2221 tggccaagct ggagaagtac tacatgaaga acggtgttga cttaggggac ccaaagaaga 2281 agcgcaaagt ggatcctaaa aagaaaagaa aggtaggcgg ccgcggggga ggcggttccg 2341 gtggcggcgg aagcggaggt ggaggatcag ggccggccat ctttaaaccc gaggagctac 2401 gccaggcact tatgccaact ctagaagcac tctatcgaca ggacccagag tctttgcctt 2461 ttcgtcagcc tgtagatcct cagctcctag gaatcccaga ttattttgat atagtgaaga 2521 atcctatgga cctttctacc atcaaacgaa agctggacac agggcaatat caagaaccct 2581 ggcagtatgt ggatgatgtc tggcttatgt tcaacaatgc gtggctatat aatcgtaaaa 2641 cgtcccgtgt atataaattt tgcagtaaac ttgcagaggt ctttgaacaa gaaattgacc 2701 ctgtcatgca gtctcttgga tattgctgtg gacgaaagta tgagttctcc ccacagactt 2761 tgtgctgtta cggaaagcag ctgtgtacaa ttcctcgtga tgcagcctac tacagctatc 2821 agaataggta tcatttctgt gagaagtgtt tcacagagat ccagggcgag aatgtgaccc 2881 tgggtgacga cccttcccaa cctcagacga caatttccaa ggatcaattt gaaaagaaga 2941 aaaatgatac cttagatcct gaaccttttg ttgactgcaa agagtgtggc cggaagatgc 3001 atcagatttg tgttctacac tatgacatca tttggccttc aggttttgtg tgtgacaact 3061 gtttgaagaa aactggcaga cctcggaaag aaaacaaatt cagtgctaag aggctgcaga 3121 ccacacgatt gggaaaccac ttagaagaca gagtgaataa gtttttgcgg cgccagaatc 3181 accctgaagc tggggaggtt tttgtcagag tggtggccag ctcagacaag actgtggagg 3241 tcaagccggg aatgaagtca aggtttgtgg attctggaga gatgtcggaa tctttcccat 3301 atcgtaccaa agcactcttt gcttttgagg agatcgatgg agtcgatgtg tgcttttttg 3361 ggatgcatgt gcaagaatac ggctctgatt gccccccacc aaatacaagg cgtgtataca 3421 tatcttatct ggacagtatt catttcttcc ggccccgctg cctccggaca gctgtttacc 3481 atgagatcct catcggatat ctcgagtatg tgaagaaatt ggggtatgtg acaggacata 3541 tttgggcctg tcccccaagt gaaggagatg actatatctt tcattgccac ccccctgacc 3601 agaaaatccc caaaccaaaa cgactacagg agtggtacaa gaagatgctg gacaaggcgt 3661 ttgcagagag gatcattaac gactataagg acatcttcaa acaagcgaac gaagacaggc 3721 tcacgagtgc caaggagttg ccctattttg aaggagattt ctggcctaat gtgttggaag 3781 aaagcattaa ggaactagaa caagaagaag aagaaaggaa aaaagaagag agtactgcag 3841 cgagtgagac tcctgagggc agtcagggtg acagcaaaaa tgcgaagaaa aagaacaaca 3901 agaagaccaa caaaaacaaa agcagcatta gccgcgccaa caagaagaag cccagcatgc 3961 ccaatgtttc caacgacctg tcgcagaagc tgtatgccac catggagaag cacaaggagg 4021 tattctttgt gattcatctg catgctgggc ctgttatcag cactcagccc cccatcgtgg 4081 accctgatcc tctgcttagc tgtgacctca tggatgggcg agatgccttc ctcaccctgg 4141 ccagagacaa gcactgggaa ttctcttcct tacgccgctc caaatggtcc actctgtgca 4201 tgctggtgga gctgcacaca cagggccagg accgctttgt ttatacctgc aatgagtgca 4261 aacaccatgt ggaaacacgc tggcactgca ctgtgtgtga ggactatgac ctttgtatca 4321 attgctacaa cacaaagagc cacacccata agatggtgaa gtgggggcta ggcctagatg 4381 atgagggcag cagtcagggt gagccacagt ccaagagccc ccaggaatcc cggcgtctca 4441 gcatccagcg ctgcatccag tccctggtgc atgcctgcca gtgtcgcaat gccaactgct 4501 cactgccgtc ttgccagaag atgaagcgag tcgtgcagca caccaagggc tgcaagcgca 4561 agactaatgg aggatgccca gtgtgcaagc agctcattgc tctttgctgc taccacgcca 4621 aacactgcca agaaaataaa tgccctgtgc ccttctgcct caacatctag ttaattaaga 4681 attcgaccca gctttcttgt acaaagtggt tgatatccag cacagtggcg gccgctcgag 4741 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc 4801 tacgcgtacc ggttagcaat tgttttttcg atgagtttgg acaaaccaca actagaatgc 4861 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 4921 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 4981 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt acttaagagg 5041 gggagaccaa agggcgagac gttaaggcct cacgtgacat gtgagcaaaa ggccagcaaa 5101 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 5161 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 5221 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 5281 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 5341 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 5401 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 5461 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 5521 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga 5581 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 5641 cttgatccgg caaacaaacc acgctggtag cggtggtttt tttgtttgca agcagcagat 5701 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 5761 tcagtggaac gaaaactcac gttaagggat tttggtcatg ccgtctcaga agaactcgtc 5821 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag 5881 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat 5941 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc 6001 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc 6061 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc 6121 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat 6181 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg 6241 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc 6301 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag 6361 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg 6421 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc 6481 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc 6541 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat 6601 aatattattg aagcatttat cagggttcgt ctcgtcccgg tctcctccca tgcatg //