LOCUS Exported 6676 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre dependent AAV expression of glutamate sensor from human synapsin promoter. ACCESSION . VERSION . KEYWORDS pAAV.hSyn.Flex.iGluSnFr.WPRE.SV40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6676) AUTHORS Marvin JS, Borghuis BG, Tian L, Cichon J, Harnett MT, Akerboom J, Gordus A, Renninger SL, Chen TW, Bargmann CI, Orger MB, Schreiter ER, Demb JB, Gan WB, Hires SA, Looger LL TITLE An optimized fluorescent probe for visualizing glutamate neurotransmission. JOURNAL Nat Methods. 2013 Feb;10(2):162-70. doi: 10.1038/nmeth.2333. Epub 2013 Jan 13. PUBMED 23314171 REFERENCE 2 (bases 1 to 6676) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6676 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 4..133 /label=AAV2 ITR intron 726..858 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 841..860 /label=pMT2-F /note="Synthetic intron, forward primer" protein_bind complement(871..904) /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(967..1000) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(1028..1174) /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" CDS complement(1178..1207) /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind 1413..1432 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1672..1693 /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1799..1820) /label=EGFP-C /note="EGFP, forward primer" sig_peptide complement(2786..2848) /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader regulatory 2845..2854 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 2879..2912 /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 2983..3016 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3088..3676 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3141..3161) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3559..3570) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3754..3875 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3765..3784) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3819..3838 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4037..4166 /label=AAV2 ITR primer_bind complement(4232..4249) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4403..4991) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4483..4502) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5162..6022) /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 5785..5804 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6023..6127) /gene="bla" /label=AmpR promoter rep_origin complement(6153..6608) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6290..6311) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6502..6521 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc 61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc 121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc 181 tctaggaaga tccgagctcc agtgtgctgg aattcgccct tttcagtatt taaattgacg 241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta 301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca 361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa 421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc 481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac 541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc 601 agcggaggag tcgtgtcgtg cctgagagcg cagctgtgct cctgggcacc gcgcagtccg 661 cccccgcggc tcctggccag accaccccta ggaccccctg ccccaagtcg cagccgttgg 721 atcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt 781 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact 841 ttgcctttct ctccacaggt gatgatcccc ataacttcgt ataaagtatc ctatacgaag 901 ttatatcaaa ataggaagac caatgcttca ccatcgaccc gaattgccaa gcatcaccat 961 cgacacataa cttcgtataa tgtatgctat acgaagttat actagtgcta gcggatccga 1021 tatcctaacg tggcttcttc tgccaaagca tgatgaggat gataagggag atgatggtga 1081 gcaccaccag ggccaggatg gctgagatca ccaccacctt aaagggcaag gagtgtggca 1141 ccacgatgac ctcctgcgtg tcctggccca cagcattcag atcctcttct gagatgagtt 1201 tttgttcgtc gacctgcagt ttcagtgcct tgtcattcgg ttctttgaac agtgctttca 1261 tttcgtctga cagttcgaaa ttcatgttca gggggttgtt aaagttgtac tccagcttat 1321 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca 1381 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga 1441 agatggtgcg ctcctggatg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct 1501 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg 1561 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt 1621 aggtggcatc gccctcaccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt 1681 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgc 1741 tccctccggt accgcccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca 1801 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttcgaa agtttggact 1861 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct 1921 gctggtagtg gtaggcgagc tgcacgccgc cgtcctcgat gttgtggcgg atcttgaagt 1981 tcgccttgat gccgttcctc tgcttgtcgg ccatgatata gacgttgtgg ctcaccagaa 2041 ttggattttt gaaccactta tcaaaccatt tttccgcttc accggaggtc tgcacctgag 2101 cgatggtgtc atccatcagc tttttgaact gcggatcatc tttacgcaac atacaaccgt 2161 aggcctcctg agactgcggc ttgccgacga tttcccagtt gtctggtttc ttcgctttcg 2221 cacgttcacc ggccagcagc acgtcatcca tcataaaggc aacggcacga ccgctttcca 2281 gggtgcggaa agagtcaccg tgatctttgg cgctgatgat gcgcatattc attttttgct 2341 cttcattcag tttgttgagc aaaacttcag aggtagtgcc ggaagtgacg actacggctt 2401 tgtctttcag gttggcaaaa tctttgatat cgccaccctt tttggtcaac aggcgcgtac 2461 cgaccacgaa aatagtgtca gagaaagccg cctgtttttg gcgttcgacg ttgttggtgg 2521 tagaaccaca ttcaaaatcg aaagtgccgt tttgcagcag tggaatacgg ttttgtgagg 2581 taatcggaat cagttttacc tgcaagtccg gtttgttgag tttctttttc actgcttcaa 2641 caatggcgtt ggagtaatcc tgcgagtaac ccaccacttt ttgctgattg tcgtaataag 2701 agaaaggcac tgaagattca cggtgaccga cgacaatcac accgtttttg gcgattttgt 2761 ccagcgtgct gcctgcggca gatctgtcac cagtggaacc tggaacccag agcagcagta 2821 cccatagcag gagtgtgtct gtctccatgg tggcgatatc accggtggta ccactagtat 2881 aacttcgtat aggatacttt atacgaagtt atcattggga ttcttcctat tttgatccaa 2941 gcatcaccat cgaccctcta gtccagatct caccatcgac ccataacttc gtatagcata 3001 cattatacga agttatgtcc ctcgaagagg ttcgaaacac tgaaaagggc gaattctgca 3061 gatgatccac tcgagaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga 3121 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 3181 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 3241 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 3301 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 3361 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 3421 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 3481 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 3541 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 3601 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt 3661 tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgctcgagca tgcatctaga 3721 gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa 3781 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt 3841 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta 3901 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat 3961 gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta gcatggcggg 4021 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct 4081 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg 4141 gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag 4201 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 4261 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 4321 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 4441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 4561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 4621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 4681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 4741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 4801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 4861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 4921 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 4981 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 5041 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 5101 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 5161 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 5221 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 5281 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 5341 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 5401 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 5461 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 5521 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 5581 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 5641 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 5701 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 5761 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 5821 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 5881 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 5941 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 6001 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 6061 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 6121 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa 6181 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa 6241 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa 6301 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca 6361 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg 6421 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc 6481 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc 6541 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca 6601 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc 6661 ctcttcgcta ttacgc //