LOCUS Exported 6199 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Medium affinity yellow glutamate sensor. ACCESSION . VERSION . KEYWORDS pAAV.hSynapsin.SF-Venus-iGluSnFR.A184V SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6199) AUTHORS Marvin JS, Scholl B, Wilson DE, Podgorski K, Kazemipour A, Muller JA, Schoch S, Quiroz FJU, Rebola N, Bao H, Little JP, Tkachuk AN, Cai E, Hantman AW, Wang SS, DePiero VJ, Borghuis BG, Chapman ER, Dietrich D, DiGregorio DA, Fitzpatrick D, Looger LL TITLE Stability, affinity, and chromatic variants of the glutamate sensor iGluSnFR. JOURNAL Nat Methods. 2018 Nov;15(11):936-939. doi: 10.1038/s41592-018-0171-3. Epub 2018 Oct 30. PUBMED 30377363 REFERENCE 2 (bases 1 to 6199) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6199 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 55..184 /label=AAV2 ITR promoter 266..713 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 742..751 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide 748..810 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader primer_bind 1776..1797 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1900..1921) /label=EGFP-N /note="EGFP, reverse primer" CDS 2386..2415 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2419..2565 /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" misc_feature 2581..3169 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2634..2654) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3052..3063) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3172..3188) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3173..3189) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 3218..3339 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3229..3248) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3283..3302 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3501..3630 /label=AAV2 ITR rep_origin complement(3844..4432) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3924..3943) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4603..5463) /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 5226..5245 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5464..5568) /gene="bla" /label=AmpR promoter rep_origin complement(5594..6049) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5731..5752) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5943..5962 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6171..6188 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcgc 61 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc 121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt 181 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag 241 atctctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg 301 gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca acccccattc 361 cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt 421 gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc 481 accgccgcct cagcactgaa ggcgcgctga cgtcactcgc tggtcccccg caaactcccc 541 ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga 601 ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc 661 tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt 721 caagctgcta gcaaggatcc cgccaccatg gagacagaca cactcctgct atgggtactg 781 ctgctctggg ttccaggttc cactggtgac agatctgccg caggcagcac gctggacaaa 841 atcgccaaaa acggtgtgat tgtcgtcggt caccgtgaat cttcagtgcc tttctcttat 901 tacgacaatc agcaaaaagt ggtgggttac tcgcaggatt actccaacgc cattgttgaa 961 gcagtgaaaa agaaactcaa caaaccggac ttgcaggtaa aactgattcc gattacctca 1021 caaaaccgta ttccactgct gcaaaacggc actttcgatt ttgaatgtgg ttctaccacc 1081 aacaacgtcg aacgccaaaa acaggcggct ttctctgaca ctattttcgt ggtcggtacg 1141 cgcctgttga ccaaaaaggg tggcgatatc aaagattttg ccaacctgaa agacaaagcc 1201 gtagtcgtca cttccggcac tacctctgaa gttttgctca acaaactgaa tgaagagcaa 1261 aaaatgaata tgcgcatcat cagcgccaaa gatcacggtg actctttccg caccctggaa 1321 agcggtcgtg ccgttgcctt tatgatggat gacgtgctgc tggccggtga acgtgcgaaa 1381 gcgaagaaac cagacaactg ggaaatcgtc ggcaagccgc agtctcagga ggcctacggt 1441 tgtatgttgc gtaaagatga tccgcagttc aaaaagctga tggatgacac catcgctcag 1501 gtgcagacct ccggtgaagc ggaaaaatgg tttgataagt ggttcaaaaa tccaattctg 1561 gtgagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggcgaacttc 1621 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac 1681 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctatcagtcc 1741 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc 1801 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg 1861 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 1921 gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag 1981 ctgaccctga agctcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 2041 accaccctgg gctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac 2101 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cagcttcaag 2161 gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 2221 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 2281 gagtacaact ttaacaaccc cctgaacatg aatttcgaac tgtcagacga aatgaaagca 2341 ctgttcaaag aaccgaatga caaggcactg aaactgcagg tcgacgaaca aaaactcatc 2401 tcagaagagg atctgaatgc tgtgggccag gacacgcagg aggtcatcgt ggtgccacac 2461 tccttgccct ttaaggtggt ggtgatctca gccatcctgg ccctggtggt gctcaccatc 2521 atctccctta tcatcctcat catgctttgg cagaagaagc cacgttagaa gcttatcgat 2581 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 2641 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 2701 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 2761 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 2821 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 2881 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 2941 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc 3001 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 3061 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 3121 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt 3181 cgacctcgac ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga 3241 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt 3301 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat 3361 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac 3421 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg 3481 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg 3541 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg 3601 ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga 3661 tcgcccttcc caacagttgc gcagcctgmm ntggcgmmkg gcwykssccs kgcsgcctca 3721 gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 3781 caacagttga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 3841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 3901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 3961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 4141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 4201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 4261 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 4321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 4381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 4441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 4501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 4561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 4621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 4681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 4741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 4801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 4861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 4921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 4981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 5101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 5161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 5221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 5281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 5341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 5401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 5461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 5521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 5581 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt 5641 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa 5701 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag 5761 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 5821 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac 5881 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag 5941 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 6001 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca 6061 ttcaggctgc gcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 6121 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 6181 tgagcgagga agcggaaga //