LOCUS       pAAV-hSyn-DIO-3xHA-hM3D(Gq)-P2A-ArgiNLS-oScarlet        6845 bp    DNA     circular UNA 16-APR-2024
DEFINITION  In the presence of Cre, the plasmid can be used to express GCaMP6s
            fused to the presynaptic protein synaptophysin. Drives enriched
            expression of GCaMP6s in neuronal axon boutons..
ACCESSION   urn.local...1ka-h40gi50
VERSION     urn.local...1ka-h40gi50
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     ##SnapGene_Notes-START##
            Confirmed Experimentally :: 0
            Transformed Into         :: unspecified
            Sequence Document Type   :: UNA
            SnapGene UUID            :: 7dc45b2b-8e0a-438f-bbf0-dc536588ebd9
            Use Custom Sequence Name :: 1
            Sequence Type            :: Synthetic
            ##SnapGene_Notes-END##
            ##Imported_From-START##
            Path     :: /Users/ericszelenyi/Downloads
            Filename :: addgene-plasmid-105715-sequence-207274.dna
            ##Imported_From-END##
FEATURES             Location/Qualifiers
     repeat_region   1..130
                     /note="Functional equivalent of wild-type AAV2 ITR"
                     /label="AAV2 ITR (alternate)"
     promoter        145..592
                     /note="human synapsin I promoter; confers neuron-specific
                     expression (<a
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/12595892""
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/12595892"">Kügl
                     er et al., 2003</a>)"
                     /label="hSyn promoter"
     protein_bind    628..661
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GGATACTT). <i>lox2272</i> sites are compatible
                     with each other, but incompatible with <i>loxP</i> or
                     <i>loxN</i> sites (<a
                     href=""https://www.ncbi.nlm.nih.gov/pubmed/9714735"">Lee
                     and Saito, 1988</a>)."
                     /bound_moiety="Cre recombinase"
                     /label="lox2272"
     protein_bind    712..745
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
                     /bound_moiety="Cre recombinase"
                     /label="loxP"
     CDS             complement(754..1524)
                     /created_by="ericszelenyi"
                     /label="ArgiNLS-oScarlet"
     regulatory      complement(1525..1590)
                     /created_by="ericszelenyi"
                     /label="P2A peptide"
     CDS             complement(1591..3378)
                     /created_by="ericszelenyi"
                     /label="hM3D(Gq)"
     Tag             complement(3379..3477)
                     /created_by="ericszelenyi"
                     /label="3x HA"
     protein_bind    complement(3493..3526)
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GGATACTT). <i>lox2272</i> sites are compatible
                     with each other, but incompatible with <i>loxP</i> or
                     <i>loxN</i> sites (<a
                     href=""https://www.ncbi.nlm.nih.gov/pubmed/9714735"">Lee
                     and Saito, 1988</a>)."
                     /bound_moiety="Cre recombinase"
                     /label="lox2272"
     protein_bind    complement(3577..3610)
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
                     /bound_moiety="Cre recombinase"
                     /label="loxP"
     misc_feature    3651..3895
                     /created_by="ericszelenyi"
                     /label="WPRE3"
     primer_bind     complement(3707..3727)
                     /Tm=55
                     /Sequence="CATAGCGTAAAAGGAGCAACA"
                     /Description="WPRE, reverse primer"
                     /label="WPRE-R"
     repeat_region   4109..4249
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /rpt_type="inverted"
                     /label="AAV2 ITR"
     repeat_region   4109..4238
                     /label="AAV2 ITR"
     rep_origin      4324..4779
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /snapgene_direction="RIGHT"
                     /label="f1 ori"
     primer_bind     complement(4411..4430)
                     /Tm=57
                     /Sequence="AGGGAAGAAAGCGAAAGGAG"
                     /Description="F1 origin, reverse primer"
                     /label="F1ori-R"
     primer_bind     4621..4642
                     /Tm=57
                     /Sequence="GTGGACTCTTGTTCCAAACTGG"
                     /Description="F1 origin, forward primer"
                     /label="F1ori-F"
     primer_bind     complement(4796..4815)
                     /Tm=54
                     /Sequence="CGGCATCAGAGCAGATTGTA"
                     /Description="pRS vectors, use to sequence yeast
                     selectable marker"
                     /label="pRS-marker"
     primer_bind     4915..4937
                     /Tm=66
                     /Sequence="CCGGGAGCTGCATGTGTCAGAGG"
                     /Description="pGEX vectors, reverse primer"
                     /label="pGEX 3'"
     primer_bind     complement(4975..4993)
                     /Tm=57
                     /Sequence="AATAGGCGTATCACGAGGC"
                     /Description="pBR322 vectors, upsteam of EcoRI site,
                     forward primer"
                     /label="pBRforEco"
     promoter        5061..5165
                     /gene="<i>bla</i>"
                     /label="AmpR promoter"
     CDS             join(5166..5234,5235..6026)
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /transl_table=1
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /product="β-lactamase"
                     /gene="<i>bla</i>"
                     /label="AmpR"
     primer_bind     complement(5384..5403)
                     /Tm=58
                     /Sequence="ATAATACCGCGCCACATAGC"
                     /Description="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     rep_origin      6197..6784
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /snapgene_direction="RIGHT"
                     /label="ori"
     primer_bind     6685..6704
                     /Tm=56
                     /Sequence="GGGAAACGCCTGGTATCTTT"
                     /Description="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
ORIGIN      
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtagtgca agtgggtttt aggaccagga tgaggcgggg
      181 tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc
      241 ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg
      301 cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca
      361 ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct
      421 tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag
      481 gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct
      541 gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agggtaccgt
      601 accggatcct ctagagtcga ctccggaata acttcgtata ggatacttta tacgaagtta
      661 tgcagaatgg tagctggatt gtagctgcta ttagcaatat gaaacctctt aataacttcg
      721 tatagcatac attatacgaa gttatggcgc gcctcacttg tacagctcgt ccatgccgcc
      781 ggtggagtgg cggccctcgg agcgttcgta ctgttccacc acggtgtagt cctcgttgtg
      841 ggaggtgatg tccaacttgc ggtccacgtt gtaggcgccg ggcatctgca cgggcttctt
      901 ggccttgtag gtggtcttga agtccgccag gtagcgcccg ccgtccttca ggcgcagggc
      961 catcttaatg tcgcccttca gcacgccgtc ctcggggtac aaccgctcgg tggacgcttc
     1021 ccagcccatt gtcttcttct gcattacggg gccgtcagga gggaagttgg tgccgcggag
     1081 cttcaccttg tagatcaggg tgccgtcctc cagggaggtg tcctgggtca cggtcacggc
     1141 gccgccgtcc tcgaagttca tcacgcggtc ccacttgaag ccctcgggga aggactgctt
     1201 atagtagtcg gggatgtcgg cggggtgctt ggtgaaggcc ctggagccgt acatgaactg
     1261 aggggacagg atgtcccagg agaagggcag ggggccaccc ttggtcacct tcagcttggc
     1321 ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga actcgtggcc
     1381 gttcatggag ccctccatgt gcaccttgaa ccgcatgaac tccttgatca ctgcctcgcc
     1441 cttgctcact cgacggcgcc gccgccttct caccttcttt ttctttgcta tgctcatctg
     1501 cttctgggga gccacccttt tcataggacc ggggttttct tccacgtctc ctgcttgctt
     1561 taacagagag aagttcgtgg ctccggatcc ggtggcgacc gggggatcct tcaaggcctg
     1621 ctcgggtgcg cgcttgtgaa aaatgaccga ctgtctctgc tggtactgct gcttgcgcct
     1681 cttttttttg tcacactggc acagcagcag catcttgaaa gtggttctga atgttttgtt
     1741 gcacagagca tagcacacgg ggttcacggt gctgttgatg tagcacagcc agtagcccag
     1801 attccaaaag gttttgggta tgcagctgtc acaaaaggtg ttcaccagaa ccatgatgtt
     1861 gtatggggtc caagtgatga tgaaggcaag caagatcgca ctgagggtct gggccgcttt
     1921 cttctccttg accagggaca tccttttccg cttagtgatc tgacttctgg tcttcagagc
     1981 aaacctcttg gccagagtgg cttccttgaa ggacagaggt agagtggccg tgctcttacc
     2041 cactgaggag ttgacgtcag aagtcttagc tgtgtccacg gctgactcta gctggatggg
     2101 aagcttggag aagctttttg gaaaactgcc tccatcgtcc acgctcttct gggcctgcag
     2161 cttgtcggct ttcctctcca agtccaccat ccccagctcc tcctcaggca cctgcaggtt
     2221 gtccgatgag ggtaacttgg tggagttgag gatggtgctg tgacccggaa gcttgagcac
     2281 gatggagtag atggctctcg tctcggagcc aatgtcctcc tcgtcggagg aggcggagtt
     2341 ctccagggag gcagcagcat cattgttgtt ccaactgtca ctgctgctgt ggtcttggtc
     2401 catctgctcg gagctgggtt tccagctctt ggttgtgaac cagaagtggc agcggccata
     2461 cttcctcctg ttggagcgtt tcatgctttg ctgttgaagt tcgtaactgc tgcagcttcg
     2521 agaactgccc gtggggtgga caaagttttc tgtctctgcc tctgtcccag aggcttgcag
     2581 gccagcaagc tctttggtac gcttttcagt ttccttatag atcctccagt ataaaatagt
     2641 cataatggtg acaggcatat aaaaaccagc gatggctgtg ccaaaagtaa tggtgggctc
     2701 actgaggaac tgaatgaagc actctcccgg aggcacagtt ctctttccaa caaagtattg
     2761 ccagaacaag atggcaggag cccaaaggac aaaggagatg acccaagcca gaccgatcat
     2821 cacaccggct ctctttgttg ttcgtttggc tcggtacgtg agcggcctcg tgatggaaaa
     2881 gtatctgtca aagctgatga ccagaagatt cataacagag gcattgctgg ctacgcagtc
     2941 aatggcaagc cagaggtcac aggccaagtt ccctaaggcc catcgattca tgatgatgta
     3001 ggtcgtaaac agattcattg aaatgacccc gataatcaga tcggcacagg ccaggcttaa
     3061 gaggaagtag ttgttgaccg tcttcagctg cttgttgacc ttaaatgaca caattaccag
     3121 gatgttgccg atgatggtca ccaaggccag gatgcccgtt aagaaagcga tgaagaccac
     3181 ttgccagacg gtatgacctc ccagagggtc atcggtggta ccgtctggag aggagaaatt
     3241 gccagctgct cgagaaacat tgtagctgcc gaaatgagtg acggttcccg ggggcagccc
     3301 tgcatcggag gggctgtgta tccaggagga gctgatgttt ggaaacaaag gcgaggttgt
     3361 actgttattg tgcaaggtag cgtagtctgg gacgtcgtat gggtagccag cgtagtctgg
     3421 gacgtcgtat gggtagccag cgtaatccgg aacatcatac gggtatcctt gttgccccat
     3481 ggtggcgcta gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag
     3541 aaattatcac tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat
     3601 acgaagttat gaattcgata tcaagcttat cgatatcgaa ttccgctcga gataatcaac
     3661 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3721 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3781 tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg
     3841 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3901 ttatttgtga aatttgtgat gctattgctt tatttgtaac catctagctt tatttgtgaa
     3961 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4021 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     4081 ggccgccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4141 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4201 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4261 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4321 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4381 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4441 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4501 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4561 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4621 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4681 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4741 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4801 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4861 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4921 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4981 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5041 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5101 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5161 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5221 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5281 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5341 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5401 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5461 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5521 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5581 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5641 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5701 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5761 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5821 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5881 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5941 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6001 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6061 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg ggttcgtgca cacagcccag
     6541 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6601 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6661 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6721 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6781 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6841 catgt
//
