LOCUS Exported 5993 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV packaging vector that expresses ER-retained GCaMP3 under control of the SYN1 promoter.. ACCESSION . VERSION . KEYWORDS pOTTC813 - pAAV-SYN1-CRTsigpep-GCaMP3-KDEL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5993) AUTHORS Henderson MJ, Baldwin HA, Werley CA, Boccardo S, Whitaker LR, Yan X, Holt GT, Schreiter ER, Looger LL, Cohen AE, Kim DS, Harvey BK TITLE A Low Affinity GCaMP3 Variant (GCaMPer) for Imaging the Endoplasmic Reticulum Calcium Store. JOURNAL PLoS One. 2015 Oct 9;10(10):e0139273. doi: 10.1371/journal.pone.0139273. eCollection 2015. PUBMED 26451944 REFERENCE 2 (bases 1 to 5993) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5993 /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 186..633 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 723..740 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 744..776 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 780..803 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" regulatory 810..819 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 831..887 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" primer_bind 1100..1121 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1227..1248) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1488..1507) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 1623..2063 /codon_start=1 /product="calcium-binding protein" /label=calmodulin /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK" misc_feature 2120..2708 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2173..2193) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2591..2602) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2740..3216 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2939..2958) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3256..3396 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3256..3385 /label=AAV2 ITR rep_origin 3471..3926 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3558..3577) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3768..3789 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3943..3962) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4062..4084 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4122..4140) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4208..4312 /gene="bla" /label=AmpR promoter CDS 4313..5173 /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(4531..4550) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5344..5932 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5833..5852 /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 cgcgtgtgtc tagactgcag agggccctgc 181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg 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 cagtcgagaa ggtaccggat ccgctagcca 661 ccatgctgct atccgtgccg ttgctgctcg gcctcctcgg cctggccgtc gccatgggtt 721 ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg 781 atctgtacga cgatgacgat aaggatctcg ccaccatggt cgactcatca cgtcgtaagt 841 ggaataagac aggtcacgca gtcagagcta taggtcggct gagctcactc gagaacgtct 901 atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca 961 tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg 1021 gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc 1081 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 1141 tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgagc aagggcgagg 1201 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 1261 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt 1321 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct 1381 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 1441 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact 1501 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 1561 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacacgc 1621 gtgaccaact gactgaagag cagatcgcag aatttaaaga ggctttctcc ctatttgaca 1681 aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctggggc 1741 agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg 1801 gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa gacacagaca 1861 gtgaagaaga aattagagaa gcgttccgtg tgtttgataa ggatggcaat ggctacatca 1921 gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg 1981 ttgatgaaat gatcagggaa gcagacatcg atggggatgg tcaggtaaac tacgaagagt 2041 ttgtacaaat gatgacagcg aagacagcag aaaaagatga gttgtagggc gcgccgaatt 2101 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg 2161 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 2221 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 2281 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 2341 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 2401 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 2461 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 2521 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg 2581 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 2641 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 2701 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc 2761 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta 2821 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg 2881 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc 2941 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc 3001 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg 3061 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg 3121 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga 3181 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc 3241 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc 3301 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg 3361 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc 3421 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg 3481 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 3541 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 3601 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 3661 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg 3721 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 3781 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt 3841 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt 3901 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc 3961 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg 4021 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca 4081 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt 4141 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg 4201 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 4261 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 4321 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 4381 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 4441 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 4501 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 4561 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 4621 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 4681 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 4741 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 4801 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 4861 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 4921 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 4981 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 5041 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 5101 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 5161 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 5221 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 5281 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 5341 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 5401 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 5461 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 5521 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 5581 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 5641 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 5701 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 5761 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 5821 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 5881 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 5941 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt //