LOCUS Exported 7104 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Adeno-associated virus plasmid, Cre-dependent FLEX switch, erHRP(N175S mutant). ACCESSION . VERSION . KEYWORDS AAV-CAG-DIO-erHRP(N175S mutant) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7104) AUTHORS Joesch M, Mankus D, Yamagata M, Shahbazi A, Shalek R, Suissa-Peleg A, Meister M, Lichtman JW, Scheirer WJ, Sanes JR TITLE Reconstruction of genetically identified neurons imaged by serial-section electron microscopy. JOURNAL Elife. 2016 Jul 7;5. pii: e15015. doi: 10.7554/eLife.15015. PUBMED 27383271 REFERENCE 2 (bases 1 to 7104) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7104 /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 enhancer 156..535 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 537..814 /label=chicken beta-actin promoter intron 816..1833 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1757..1779 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1841..1860 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 1912..1945 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 1996..2029 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(2986..3012) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" sig_peptide complement(3013..3075) /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader regulatory 3072..3081 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(3089..3122) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(3173..3206) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3231..3819 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3284..3304) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3702..3713) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3851..4327 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4050..4069) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4367..4507 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4367..4496 /label=AAV2 ITR rep_origin 4582..5037 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4669..4688) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4879..4900 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5054..5073) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5173..5195 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5233..5251) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5319..5423 /gene="bla" /label=AmpR promoter CDS 5424..6284 /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(5642..5661) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6455..7043 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6944..6963 /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 cgctcgacat tgattattga ctagttatta 181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga 541 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt 601 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg 661 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc 721 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg 781 gcggccctat aaaaagcgaa gcgcgcggcg ggcggggagt cgctgcgacg ctgccttcgc 841 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta 901 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt 961 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc 1021 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc 1081 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt 1141 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg 1201 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt 1261 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac 1321 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc 1381 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc 1441 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc 1501 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc 1561 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag 1621 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc 1681 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg 1741 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct 1801 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg 1861 ctgtctcatc attttggcaa agaattcgga tcctctagag tcgactccgg aataacttcg 1921 tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct gctattagca 1981 atatgaaacc tcttaataac ttcgtatagc atacattata cgaagttatg gcgcgcctta 2041 aagttcgtcc ttcaagtcag agttgctgtt cacgacccta cagttcagtc ttatctgtcc 2101 ctgcgtaccc gtcaggggcg ttatatttcc catccgatcc atagcttcca cgaaagcatt 2161 gaaaaaggtt tgtgtgctat tagcgaagct gcgcacaaga ggaatagtgt cggtagcatt 2221 tggggaactg aacagctcct ggtcggactg aatcaggccc ttctgttcct ccagattgac 2281 gtaatactta ttgtcgaaga tggttggggt gcgcaggtca aagtcaacca gtgcagacaa 2341 gttgccattg agcggacaca gccctctgag agtctgcaag tatgttgtgt tcagggtagg 2401 gtcaggcaag ccggtgttgg aaaagttata cagtctgtcc atgatgaacc tgcactgact 2461 cttgccgaat gtgtggcctc cggacagagc caccaaatca ctacttcggt tcaggccgac 2521 attcctgaag gaatccttca gctggggaag ggtgaagaag ggagcaggca ggtttgcgtt 2581 tgccaagtcc agaaaagcct gcagagagtc cctccttccc agggggaccc tccagctggg 2641 gccaccggcc agcgtcacgg attgctgggc cgctatggtc agaaggtctg cacaactaac 2701 tgtccggggg caggcgctct ccacagcggc tttcattcta tcgatcacgg gaaaccctct 2761 agcactattc gcgttaccga aggcatcctt ttcagtcctg aaagaggtgg tattgtcgag 2821 gaggatggaa gcatcacagc cgtttacaaa acaatcgtgg aaatgcagcc gcaaaattga 2881 agcggctatc ctcggatcag accgcaactc gttcacgata gtatctcgca cgatgttgct 2941 cacgttagga caagagttat cgtagaaagt tggagtaagc tgcatagcat aatctggaac 3001 atcatatgga tagtcaccag tggaacctgg aacccagagc agcagtaccc atagcaggag 3061 tgtgtctgtc tccatggtgg cggctagcat aacttcgtat aaagtatcct atacgaagtt 3121 atttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc 3181 gtataatgta tgctatacga agttatgaat tcgatatcaa gcttatcgat aatcaacctc 3241 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc 3301 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca 3361 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg 3421 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca 3481 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg 3541 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg 3601 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg 3661 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg 3721 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc 3781 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg 3841 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt 3901 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga 3961 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt 4021 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc 4081 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc 4141 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg 4201 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat 4261 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc 4321 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg 4381 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag 4441 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc 4501 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 4561 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 4621 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt 4681 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc 4741 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg 4801 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg 4861 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct 4921 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg 4981 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat 5041 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc 5101 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag 5161 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg 5221 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg 5281 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 5341 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 5401 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct 5461 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag 5521 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta 5581 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc 5641 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca 5701 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg 5761 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg 5821 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca 5881 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa 5941 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa 6001 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata 6061 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat 6121 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc 6181 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata 6241 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt 6301 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 6361 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 6421 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 6481 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 6541 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 6601 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 6661 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 6721 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 6781 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 6841 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 6901 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 6961 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 7021 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 7081 tttgctggcc ttttgctcac atgt //