LOCUS Exported 6656 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mCBPHAT_4xNLS_PUFa_2xNLS in pmax expression vector. ACCESSION . VERSION . KEYWORDS pAC1418-pmax-mCBPHAT_4xNLS_PUFa_2xNLS 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 12, 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" regulatory 2628..2637 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3324..3344 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3348..3368 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3372..3392 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3483..3503 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4560..4580 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4584..4604 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" 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 atctttaaac 1021 ccgaggagct acgccaggca cttatgccaa ctctagaagc actctatcga caggacccag 1081 agtctttgcc ttttcgtcag cctgtagatc ctcagctcct aggaatccca gattattttg 1141 atatagtgaa gaatcctatg gacctttcta ccatcaaacg aaagctggac acagggcaat 1201 atcaagaacc ctggcagtat gtggatgatg tctggcttat gttcaacaat gcgtggctat 1261 ataatcgtaa aacgtcccgt gtatataaat tttgcagtaa acttgcagag gtctttgaac 1321 aagaaattga ccctgtcatg cagtctcttg gatattgctg tggacgaaag tatgagttct 1381 ccccacagac tttgtgctgt tacggaaagc agctgtgtac aattcctcgt gatgcagcct 1441 actacagcta tcagaatagg tatcatttct gtgagaagtg tttcacagag atccagggcg 1501 agaatgtgac cctgggtgac gacccttccc aacctcagac gacaatttcc aaggatcaat 1561 ttgaaaagaa gaaaaatgat accttagatc ctgaaccttt tgttgactgc aaagagtgtg 1621 gccggaagat gcatcagatt tgtgttctac actatgacat catttggcct tcaggttttg 1681 tgtgtgacaa ctgtttgaag aaaactggca gacctcggaa agaaaacaaa ttcagtgcta 1741 agaggctgca gaccacacga ttgggaaacc acttagaaga cagagtgaat aagtttttgc 1801 ggcgccagaa tcaccctgaa gctggggagg tttttgtcag agtggtggcc agctcagaca 1861 agactgtgga ggtcaagccg ggaatgaagt caaggtttgt ggattctgga gagatgtcgg 1921 aatctttccc atatcgtacc aaagcactct ttgcttttga ggagatcgat ggagtcgatg 1981 tgtgcttttt tgggatgcat gtgcaagaat acggctctga ttgcccccca ccaaatacaa 2041 ggcgtgtata catatcttat ctggacagta ttcatttctt ccggccccgc tgcctccgga 2101 cagctgttta ccatgagatc ctcatcggat atctcgagta tgtgaagaaa ttggggtatg 2161 tgacaggaca tatttgggcc tgtcccccaa gtgaaggaga tgactatatc tttcattgcc 2221 acccccctga ccagaaaatc cccaaaccaa aacgactaca ggagtggtac aagaagatgc 2281 tggacaaggc gtttgcagag aggatcatta acgactataa ggacatcttc aaacaagcga 2341 acgaagacag gctcacgagt gccaaggagt tgccctattt tgaaggagat ttctggccta 2401 atgtgttgga agaaagcatt aaggaactag aacaagaaga agaagaaagg aaaaaagaag 2461 agagtactgc agcgagtgag actcctgagg gcagtcaggg tgacagcaaa aatgcgaaga 2521 aaaagaacaa caagaagacc aacaaaaaca aaagcagcat tagccgcgcc aacaagaaga 2581 agcccagcat gcccaatgtt tccaacgacc tgtcgcagaa gctgtatgcc accatggaga 2641 agcacaagga ggtattcttt gtgattcatc tgcatgctgg gcctgttatc agcactcagc 2701 cccccatcgt ggaccctgat cctctgctta gctgtgacct catggatggg cgagatgcct 2761 tcctcaccct ggccagagac aagcactggg aattctcttc cttacgccgc tccaaatggt 2821 ccactctgtg catgctggtg gagctgcaca cacagggcca ggaccgcttt gtttatacct 2881 gcaatgagtg caaacaccat gtggaaacac gctggcactg cactgtgtgt gaggactatg 2941 acctttgtat caattgctac aacacaaaga gccacaccca taagatggtg aagtgggggc 3001 taggcctaga tgatgagggc agcagtcagg gtgagccaca gtccaagagc ccccaggaat 3061 cccggcgtct cagcatccag cgctgcatcc agtccctggt gcatgcctgc cagtgtcgca 3121 atgccaactg ctcactgccg tcttgccaga agatgaagcg agtcgtgcag cacaccaagg 3181 gctgcaagcg caagactaat ggaggatgcc cagtgtgcaa gcagctcatt gctctttgct 3241 gctaccacgc caaacactgc caagaaaata aatgccctgt gcccttctgc ctcaacatca 3301 acgatggcgg aggcggatct gatccaaaaa agaagagaaa ggtagatcca aaaaagaaga 3361 gaaaggtaga tccaaaaaag aagagaaagg taggatctac cggatctaga aacgatggtg 3421 gtggtggaag cgggggtggg ggcagcggtg gagggggaag cgggcgcgcc gggatcctcc 3481 cccccaagaa aaagaggaag gtatctagag gccgcagccg ccttttggaa gattttcgaa 3541 acaaccggta ccccaattta caactgcggg agattgctgg acatataatg gaattttccc 3601 aagaccagca tgggtccaga ttcattcagc tgaaactgga gcgtgccaca ccagctgagc 3661 gccagcttgt cttcaatgaa atcctccagg ctgcctacca actcatggtg gatgtgtttg 3721 gtaattacgt cattcagaag ttctttgaat ttggcagtct tgaacagaag ctggctttgg 3781 cagaacggat tcgaggccac gtcctgtcat tggcactaca gatgtatggc agccgtgtta 3841 tcgagaaagc tcttgagttt attccttcag accagcagaa tgagatggtt cgggaactag 3901 atggccatgt cttgaagtgt gtgaaagatc agaatggcaa tcacgtggtt cagaaatgca 3961 ttgaatgtgt acagccccag tctttgcaat ttatcatcga cgcgtttaag ggacaggtat 4021 ttgccttatc cacacatcct tatggctgcc gagtgattca gagaatcctg gagcactgtc 4081 tccctgacca gacactccct attttagagg agcttcacca gcacacagag cagctggtac 4141 aggatcaata tggaaattat gtaatccaac atgtactgga gcacggtcgt cctgaggata 4201 aaagcaaaat tgtagcagaa atccgaggca atgtacttgt attgagtcag cacaaatttg 4261 caagcaatgt tgtggagaag tgtgttactc acgcctcacg tacggagcgc gctgtgctca 4321 tcgacgaggt gtgcaccatg aacgacggtc cccacagtgc cttatacacc atgatgaagg 4381 accagtatgc caactacgtg gtccagaaga tgattgacgt ggcggagcca ggccagcgga 4441 agatcgtcat gcataagatc cggccccaca tcgcaactct tcgtaagtac acctatggca 4501 agcacattct ggccaagctg gagaagtact acatgaagaa cggtgttgac ttaggggacc 4561 caaagaagaa gcgcaaagtg gatcctaaaa agaaaagaaa ggtaggcggc cgcgggggag 4621 gcggttccgg tggcggcgga agcggaggtg gaggatcagg gccggcctga 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 //