LOCUS pAAV-CAG-FLPo-3xmiR122-WPRE-final 6565 bp DNA circular UNA 07-MAY-2024 DEFINITION FLPo for Addgene submission. ACCESSION urn.local...1n-howdgh3 VERSION urn.local...1n-howdgh3 KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers misc_feature 1..130 /label="L-ITR" misc_feature 46..47 /note="aaagcccgggc" /label="Missing portion of the ITR" promoter 153..1252 /note="CAG" /label="CAG" source <1268..>2581 /organism="synthetic DNA construct" /mol_type="other DNA" /label="source synthetic DNA construct" misc_feature 1269..2580 /Original_Bases="CCACCATGTCTAGTGATGATGAGGCTACTGCTGACTCTCAA CATTCTACTCCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCATGGTGAGCAAGGGCG AGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGG CCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACC CTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCA CCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGA CTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAG GACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGA ACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAA GCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAAC GGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCG CCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAA CCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCAC ATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGT ACAAGTA" /label="CCACCATGTCTAGTGATGATGAGGCTACTGCTGACTCTCAACATTCTACT CCTCCAAAAAAGAAGAGAAAGGTAGAAGACCCCATGGTGAGCAAGGGCGAGGAGCTGT TCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTT CAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTC ATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCT ACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAA GTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGC AACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCG AGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTA CAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAG GTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACT ACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCT GAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTG CTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTA" /note="Geneious type: Editing History Replacement" regulatory 1277..1286 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /label="regulatory" CDS 1283..2581 /codon_start=1 /product="nuclear-targeted site-specific recombinase" /note="FLPo is a mouse codon-optimized fusion between the SV40 nuclear localization signal and a thermostable versionof the Saccharomyces cerevisiae site-specific recombinaseFLP. Recombination occurs at FRT sequences." /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCA AELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASL KKLIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEG ESIWEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNK YLGVIIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSS NKQEYQLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTN VVGNWSDKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEE WQHIEQLKGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI" /label="FLPo" misc_feature 2591..2673 /label="Inside template" misc_feature 2594..2615 /label="mmu-miR-122-5p Binding site" misc_feature 2616..2621 /label="spacer 1" misc_feature 2622..2643 /label="mmu-miR-122-5p Binding site" misc_feature 2644..2649 /label="spacer 2" misc_feature 2650..2671 /label="mmu-miR-122-5p Binding site" misc_feature 2682..3290 /label="WPRE" polyA_signal 3310..3788 /label="hGH polyA" misc_feature 3828..3968 /label="R-ITR" rep_origin 4060..4366 /direction="RIGHT" /label="f1 Origin" misc_feature 4885..5742 /label="AmpR" rep_origin 5893..6560 /direction="RIGHT" /label="pUC ori" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtaacact agttattaat agtaatcaat tacggggtca 181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 481 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc 541 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta 601 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg 661 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg 721 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc 781 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc 841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac 901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg 961 tggctgcgtg aaagccttga ggggctccgg gagcgccggc aggaaggaaa tgggcgggga 1021 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc 1081 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt 1141 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct 1201 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgcggcccaa 1261 cggtaccgga tccgccgcca ccatggctcc taagaagaag aggaaggtga tgagccagtt 1321 cgacatcctg tgcaagaccc cccccaaggt gctggtgcgg cagttcgtgg agagattcga 1381 gaggcccagc ggcgagaaga tcgccagctg tgccgccgag ctgacctacc tgtgctggat 1441 gatcacccac aacggcaccg ccatcaagag ggccaccttc atgagctaca acaccatcat 1501 cagcaacagc ctgagcttcg acatcgtgaa caagagcctg cagttcaagt acaagaccca 1561 gaaggccacc atcctggagg ccagcctgaa gaagctgatc cccgcctggg agttcaccat 1621 catcccttac aacggccaga agcaccagag cgacatcacc gacatcgtgt ccagcctgca 1681 gctgcagttc gagagcagcg aggaggccga caagggcaac agccacagca agaagatgct 1741 gaaggccctg ctgtccgagg gcgagagcat ctgggagatc accgagaaga tcctgaacag 1801 cttcgagtac accagcaggt tcaccaagac caagaccctg taccagttcc tgttcctggc 1861 cacattcatc aactgcggca ggttcagcga catcaagaac gtggacccca agagcttcaa 1921 gctggtgcag aacaagtacc tgggcgtgat cattcagtgc ctggtgaccg agaccaagac 1981 aagcgtgtcc aggcacatct actttttcag cgccagaggc aggatcgacc ccctggtgta 2041 cctggacgag ttcctgagga acagcgagcc cgtgctgaag agagtgaaca ggaccggcaa 2101 cagcagcagc aacaagcagg agtaccagct gctgaaggac aacctggtgc gcagctacaa 2161 caaggccctg aagaagaacg ccccctaccc catcttcgct atcaagaacg gccctaagag 2221 ccacatcggc aggcacctga tgaccagctt tctgagcatg aagggcctga ccgagctgac 2281 aaacgtggtg ggcaactgga gcgacaagag ggcctccgcc gtggccagga ccacctacac 2341 ccaccagatc accgccatcc ccgaccacta cttcgccctg gtgtccaggt actacgccta 2401 cgaccccatc agcaaggaga tgatcgccct gaaggacgag accaacccca tcgaggagtg 2461 gcagcacatc gagcagctga agggcagcgc cgagggcagc atcagatacc ccgcctggaa 2521 cggcatcatc agccaggagg tgctggacta cctgagcagc tacatcaaca ggcggatctg 2581 agaattcacc ggtcaaacac cattgtcaca ctccatacta tcaaacacca ttgtcacact 2641 ccagatcaac aaacaccatt gtcacactcc acgccggcga agcttatcga taatcaacct 2701 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 2761 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 2821 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 2881 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 2941 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 3001 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 3061 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt 3121 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 3181 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 3241 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc 3301 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag 3361 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg 3421 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag 3481 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg 3541 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc 3601 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt 3661 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga 3721 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc 3781 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt 3841 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa 3901 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg 3961 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg 4021 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg 4081 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct 4141 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg 4201 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg 4261 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg 4321 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc 4381 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat 4441 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta 4501 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg 4561 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa 4621 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc 4681 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg 4741 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 4801 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 4861 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 4921 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 4981 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 5041 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 5101 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc 5161 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 5221 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 5281 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 5341 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 5401 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 5461 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 5521 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 5581 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag 5641 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 5701 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 5761 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 5821 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga 5881 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 5941 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 6001 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 6061 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 6121 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 6181 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 6241 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 6301 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 6361 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 6421 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 6481 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 6541 ttttgctggc cttttgctca catgt //