LOCUS Exported 4972 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Can be used to generate AAV virus that will express FlpE recombinse gene in neurons from the synapsin promoter. ACCESSION . VERSION . KEYWORDS AAV phSyn1(S)-FlpE-bGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4972) AUTHORS Madisen L, Garner AR, Shimaoka D, Chuong AS, Klapoetke NC, Li L, van der Bourg A, Niino Y, Egolf L, Monetti C, Gu H, Mills M, Cheng A, Tasic B, Nguyen TN, Sunkin SM, Benucci A, Nagy A, Miyawaki A, Helmchen F, Empson RM, Knopfel T, Boyden ES, Reid RC, Carandini M, Zeng H TITLE Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance. JOURNAL Neuron. 2015 Mar 4;85(5):942-58. doi: 10.1016/j.neuron.2015.02.022. PUBMED 25741722 REFERENCE 2 (bases 1 to 4972) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4972 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 177..624 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 658..1956 /codon_start=1 /product="nuclear-targeted site-specific recombinase" /label=FLPe /note="FLPe is a fusion between the SV40 nuclear localization signal and a thermostable version of the Saccharomyces cerevisiae site-specific recombinase FLP. Recombination occurs at FRT sequences." /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI" primer_bind complement(1999..2016) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2005..2212 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 2235..2375 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2235..2364 /label=AAV2 ITR rep_origin 2450..2905 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2537..2556) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2747..2768 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2922..2941) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3041..3063 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3101..3119) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3187..3291 /gene="bla" /label=AmpR promoter CDS 3292..4152 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3510..3529) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4323..4911 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4812..4831 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtctgca gagggccctg cgtatgagtg 181 caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc 241 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg 301 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca 361 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg 421 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc 481 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac 541 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga 601 gtcgtgtcgt gcctgagagc gcagacgcgt ggacggtacc cttcaggtcg acccaccatg 661 gctcccaaga agaagaggaa ggtgatgagt caatttgata tattatgtaa aacaccacct 721 aaggtcctgg ttcgtcagtt tgtggaaagg tttgaaagac cttcagggga aaaaatagca 781 tcatgtgctg ctgaactaac ctatttatgt tggatgatta ctcataacgg aacagcaatc 841 aagagagcca cattcatgag ctataatact atcataagca attcgctgag tttcgatatt 901 gtcaacaaat cactccagtt taaatacaag acgcaaaaag caacaattct ggaagcctca 961 ttaaagaaat taattcctgc ttgggaattt acaattattc cttacaatgg acaaaaacat 1021 caatctgata tcactgatat tgtaagtagt ttgcaattac agttcgaatc atcggaagaa 1081 gcagataagg gaaatagcca cagtaaaaaa atgcttaaag cacttctaag tgagggtgaa 1141 agcatctggg agatcactga gaaaatacta aattcgtttg agtatacctc gagatttaca 1201 aaaacaaaaa ctttatacca attcctcttc ctagctactt tcatcaattg tggaagattc 1261 agcgatatta agaacgttga tccgaaatca tttaaattag tccaaaataa gtatctggga 1321 gtaataatcc agtgtttagt gacagagaca aagacaagcg ttagtaggca catatacttc 1381 tttagcgcaa ggggtaggat cgatccactt gtatatttgg atgaattttt gaggaattct 1441 gaaccagtcc taaaacgagt aaataggacc ggcaattctt caagcaacaa acaggaatac 1501 caattattaa aagataactt agtcagatcg tacaacaagg ctttgaagaa aaatgcgcct 1561 tatccaatct ttgctataaa gaatggccca aaatctcaca ttggaagaca tttgatgacc 1621 tcatttctgt caatgaaggg cctaacggag ttgactaatg ttgtgggaaa ttggagcgat 1681 aagcgtgctt ctgccgtggc caggacaacg tatactcatc agataacagc aatacctgat 1741 cactacttcg cactagtttc tcggtactat gcatatgatc caatatcaaa ggaaatgata 1801 gcattgaagg atgagactaa tccaattgag gagtggcagc atatagaaca gctaaagggt 1861 agtgctgaag gaagcatacg ataccccgca tggaatggga taatatcaca ggaggtacta 1921 gactaccttt catcctacat aaatagacgc atataatgat ctagagtcga gttaattaag 1981 aattccgccc cagatctgcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2041 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 2101 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 2161 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gacacgtgcg 2221 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg 2281 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc 2341 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct 2401 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt 2461 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 2521 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 2581 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 2641 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga 2701 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 2761 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg 2821 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 2881 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc 2941 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 3001 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 3061 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt 3121 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 3181 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 3241 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 3301 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 3361 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 3421 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 3481 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 3541 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 3601 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 3661 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 3721 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 3781 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 3841 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 3901 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 3961 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 4021 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 4081 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 4141 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 4201 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 4261 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 4321 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 4381 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 4441 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac 4501 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 4561 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 4621 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 4681 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 4741 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 4801 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 4861 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 4921 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt //