LOCUS Exported 8515 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION A backbone for adenovirus creation by the AdEasy method, similar to ShuttleCMV (Addgene Plasmid # 16403), except it uses a mouse cofilin promoter (MCP) instead of the CMV promoter. ACCESSION . VERSION . KEYWORDS ShuttleMCP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8515) AUTHORS Mi J, Shaw AE, Pak CW, Walsh KP, Minamide LS, Bernstein BW, Kuhn TB, Bamburg JR TITLE A genetically encoded reporter for real-time imaging of cofilin-actin rods in living neurons. JOURNAL PLoS One. 2013 Dec 31;8(12):e83609. doi: 10.1371/journal.pone.0083609. PUBMED 24391794 REFERENCE 2 (bases 1 to 8515) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8515 /organism="synthetic DNA construct" /mol_type="other DNA" misc_signal 202..352 /label=Ad5 Psi /note="packaging signal for adenovirus serotype 5" CDS complement(640..651) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" primer_bind complement(5735..5752) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5906..6494) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5986..6005) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 7307..8101 /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(7361..7380) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7971..7990 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(8211..8232) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8423..8442 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 ttaattaann ntcccttcca gctctctgcc ccttttggat tgaagccaat atgataatga 61 gggggtggag tttgtgacgt ggcgcggggc gtgggaacgg ggcgggtgac gtagtagtgt 121 ggcggaagtg tgatgttgca agtgtggcgg aacacatgta agcgacggat gtggcaaaag 181 tgacgttttt ggtgtgcgcc ggtgtacaca ggaagtgaca attttcgcgc ggttttaggc 241 ggatgttgta gtaaatttgg gcgtaaccga gtaagatttg gccattttcg cgggaaaact 301 gaataagagg aagtgaaatc tgaataattt tgtgttactc atagcgcgta atactggtac 361 cgcttcggcc tccacctgga gctccagcag tgtagcccac gggtgcagtc acctggagct 421 cccgcagtgt agcccacggg tgcagtctga ctacaatccc ctgactacaa cccccggcat 481 cctctgcgtg ctgcctcgct cctggggaag ctcctctttt acacccttta ttcagcttgg 541 ggaggtcaag ggccccggtt tcctttgcct ctgagttgtt tgcgactttg cccttttttt 601 ctaaggccat ggtcattgtt tcaggtcaga gaaattctcc tggagttccc accccagttt 661 attttaaact tttgctttca tatagattct cttcagtctt tgtgctaaaa tgatcataaa 721 cctatcaact ctggaattaa attgaagtat tccttttcct gtatcgtaaa agaaacataa 781 agaagttttt tttttgtttt gttttgtttt gttttgtttt ttgatttttc aagacagggt 841 ttctctgtgt agccctggct gtcctggaac tcactttgta gaccaggctt gcctcgcact 901 cagaaatccg cctgcctctg cctcccgagt gctgggatta aaggtgtgcg ccaccatgcc 961 cggccataag gaaattttta gtgcacaact cttcaccctt cccccatgaa ttacgtttga 1021 tctaacaatt tgccaaacgt ctttagacaa cgccaaagct ctcaaacttt ttatgttagc 1081 atttcctgtt ttttaaaaaa tcctatttgt tgaaatttta ctccacggga gcatttctat 1141 tggcttgaag acaccgttat acttggcgat tctcttttct gtgagagagg atcccaagct 1201 gatgattcaa aatctttttt caggaaccat aaagaacaaa aaaccactta atttctggga 1261 taaagaaagt agagaattat attcaggtca tttttatcaa acgatgggcc tctggctcag 1321 agcgtttttt ggtccattgc accaatgggc tcactctccg tgcaacagcc cacgcctctc 1381 ggcctccgcc tactctgcgc ctcaagctct gcggaacccg gtagcagcta gttctatttc 1441 ggtgcaaaat cctcggactt cacttcccgt cggcccgcgc ggcaagcgcc ggaaggccgc 1501 cccgcccctc attgtgcggc tcctactaaa cggaaggggc cgggagaggc cgcgttcagt 1561 cggatcccgg cagcagctgc ggcggctcag gtcttcagtt gctctttctg tctcccggta 1621 ccgcggccgc ctcgagtcta gagatatcga attcaagctt gtcgactcga agatctgccg 1681 tcatttccct ggagggcaag cctttgtgag ccacctccag ccccctgcct ggagcatctt 1741 aacagcccca gacctgctct tgggtgttgc aggctgcccc tttcctgcca gaccggaggg 1801 gctgggggga tcccagcagg gggagggcta tcccttcacc ccagttgcca aacatccctc 1861 ccaccccctg gaccgtcctt ctccctccat ccctgacggt tctggccttc ccaaactgct 1921 tttgatcttc tgattcctct tgggttgacg cagaccaagt cccgtcctag gcacccagtt 1981 tggggggagc ctgtattttt ttttttaacg acacccctac tccgtatccc tccccatccc 2041 atgctgccaa cttctaacca caatagtgac tctgtgcttg tctgtttagt tctgtgtgta 2101 aatgaaatgt ggaaatgacc ctccctgccc cagctggctg ccctcccctt tcctttgatc 2161 ttgaccactc atggaagcag gaccagtaag ggaccttcaa tttaaaacaa aacaaaacaa 2221 aaaaacaata aaaaggctaa ttaacaacat ggcttttggg tttgggattt tggggaccct 2281 gtggtgggag atggatgggt ggtttccctg tgcttcacct ctagttcagt ccctagtaag 2341 cggcatctcc ttcttgagtt ggggtgcact gttccaaggc agggcagttg ggctcagatc 2401 tgggcgtggt taagggtggg aaagaatata taaggtgggg gtcttatgta gttttgtatc 2461 tgttttgcag cagccgccgc cgccatgagc accaactcgt ttgatggaag cattgtgagc 2521 tcatatttga caacgcgcat gcccccatgg gccggggtgc gtcagaatgt gatgggctcc 2581 agcattgatg gtcgccccgt cctgcccgca aactctacta ccttgaccta cgagaccgtg 2641 tctggaacgc cgttggagac tgcagcctcc gccgccgctt cagccgctgc agccaccgcc 2701 cgcgggattg tgactgactt tgctttcctg agcccgcttg caagcagtgc agcttcccgt 2761 tcatccgccc gcgatgacaa gttgacggct cttttggcac aattggattc tttgacccgg 2821 gaacttaatg tcgtttctca gcagctgttg gatctgcgcc agcaggtttc tgccctgaag 2881 gcttcctccc ctcccaatgc ggtttaaaac ataaataaaa aaccagactc tgtttggatt 2941 tggatcaagc aagtgtcttg ctgtctttat ttaggggttt tgcgcgcgcg gtaggcccgg 3001 gaccagcggt ctcggtcgtt gagggtcctg tgtatttttt ccaggacgtg gtaaaggtga 3061 ctctggatgt tcagatacat gggcataagc ccgtctctgg ggtggaggta gcaccactgc 3121 agagcttcat gctgcggggt ggtgttgtag atgatccagt cgtagcagga gcgctgggcg 3181 tggtgcctaa aaatgtcttt cagtagcaag ctgattgcca ggggcaggcc cttggtgtaa 3241 gtgtttacaa agcggttaag ctgggatggg tgcatacgtg gggatatgag atgcatcttg 3301 gactgtattt ttaggttggc tatgttccca gccatatccc tccggggatt catgttgtgc 3361 agaaccacca gcacagtgta tccggtgcac ttgggaaatt tgtcatgtag cttagaagga 3421 aatgcgtgga agaacttgga gacgcccttg tgacctccaa gattttccat gcattcgtcc 3481 ataatgatgg caatgggccc acgggcggcg gcctgggcga agatatttct gggatcacta 3541 acgtcatagt tgtgttccag gatgagatcg tcataggcca tttttacaaa gcgcgggcgg 3601 agggtgccag actgcggtat aatggttcca tccggcccag gggcgtagtt accctcacag 3661 atttgcattt cccacgcttt gagttcagat ggggggatca tgtctacctg cggggcgatg 3721 aagaaaacgg tttccggggt aggggagatc agctgggaag aaagcaggtt cctgagcagc 3781 tgcgacttac cgcagccggt gggcccgtaa atcacaccta ttaccgggtg caactggtag 3841 ttaagagagc tgcagctgcc gtcatccctg agcagggggg ccacttcgtt aagcatgtcc 3901 ctgactcgca tgttttccct gaccaaatcc gccagaaggc gctcgccgcc cagcgatagc 3961 agttcttgca aggaagcaaa gtttttcaac ggtttgagac cgtccgccgt aggcatgctt 4021 ttgagcgttt gaccaagcag ttccaggcgg tcccacagct cggtcacctg ctctacggca 4081 tctcgatcca gcatatctcc tcgtttcgcg ggttggggcg gctttcgctg tacggcagta 4141 gtcggtgctc gtccagacgg gccagggtca tgtctttcca cgggcgcagg gtcctcgtca 4201 gcgtagtctg ggtcacggtg aaggggtgcg ctccgggctg cgcgctggcc agggtgcgct 4261 tgaggctggt cctgctggtg ctgaagcgct gccggtcttc gccctgcgcg tcggccaggt 4321 agcatttgac catggtgtca tagtccagcc cctccgcggc gtggcccttg gcgcgcagct 4381 tgcccttgga ggaggcgccg cacgaggggc agtgcagact tttgagggcg tagagcttgg 4441 gcgcgagaaa taccgattcc ggggagtagg catccgcgcc gcaggccccg cagacggtct 4501 cgcattccac gagccaggtg agctctggcc gttcggggtc aaaaaccagg tttcccccat 4561 gctttttgat gcgtttctta cctctggttt ccatgagccg gtgtccacgc tcggtgacga 4621 aaaggctgtc cgtgtccccg tatacagact nnngtttaaa cgaattcnnn tataaaatgc 4681 aaggtgctgc tcaaaaaatc aggcaaagcc tcgcgcaaaa aagaaagcac atcgtagtca 4741 tgctcatgca gataaaggca ggtaagctcc ggaaccacca cagaaaaaga caccattttt 4801 ctctcaaaca tgtctgcggg tttctgcata aacacaaaat aaaataacaa aaaaacattt 4861 aaacattaga agcctgtctt acaacaggaa aaacaaccct tataagcata agacggacta 4921 cggccatgcc ggcgtgaccg taaaaaaact ggtcaccgtg attaaaaagc accaccgaca 4981 gctcctcggt catgtccgga gtcataatgt aagactcggt aaacacatca ggttgattca 5041 tcggtcagtg ctaaaaagcg accgaaatag cccgggggaa tacatacccg caggcgtaga 5101 gacaacatta cagcccccat aggaggtata acaaaattaa taggagagaa aaacacataa 5161 acacctgaaa aaccctcctg cctaggcaaa atagcaccct cccgctccag aacaacatac 5221 agcgcttcac agcggcagcc taacagtcag ccttaccagt aaaaaagaaa acctattaaa 5281 aaaacaccac tcgacacggc accagctcaa tcagtcacag tgtaaaaaag ggccaagtgc 5341 agagcgagta tatataggac taaaaaatga cgtaacggtt aaagtccaca aaaaacaccc 5401 agaaaaccgc acgcgaacct acgcccagaa acgaaagcca aaaaacccac aacttcctca 5461 aatcgtcact tccgttttcc cacgttacgt aacttcccat tttaagaaaa ctacaattcc 5521 caacacatac aagttactcc gccctaaaac ctacgtcacc cgccccgttc ccacgccccg 5581 cgccacgtca caaactccac cccctcatta tcatattggc ttcaatccaa aataaggtat 5641 attattgatg atnnnttaat taaggatccn nncggtgtga aataccgcac agatgcgtaa 5701 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 5761 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 5821 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 5881 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 5941 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 6001 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 6061 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 6121 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 6181 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 6241 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 6301 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 6361 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 6421 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 6481 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 6541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 6601 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 6661 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 6721 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 6781 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 6841 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 6901 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 6961 gcaacgttgt tgnnnnnnaa aaaggatctt cacctagatc cttttcacgt agaaagccag 7021 tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga 7081 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga 7141 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa 7201 ggttgggaag ccctgcaaag taaactggat ggctttctcg ccgccaagga tctgatggcg 7261 caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 7321 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 7381 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 7441 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 7501 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 7561 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 7621 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 7681 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 7741 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 7801 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 7861 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 7921 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 7981 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 8041 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 8101 aattttgtta aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 8161 acatccctta taaatcaaaa gaatagaccg cgatagggtt gagtgttgtt ccagtttgga 8221 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 8281 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgcgg tcgaggtgcc 8341 gtaaagctct aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc 8401 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg 8461 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc gcgcttaatg cgccg //