LOCUS       AAV-hSyn-DIO-FLAG-hM4D(Gi)-P2A-ArgiNLS-AausFP1        6533 bp    DNA     circular UNA 19-APR-2024
DEFINITION  .
ACCESSION   urn.local...1rv-hibjbi2
VERSION     urn.local...1rv-hibjbi2
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     ##SnapGene_Notes-START##
            Confirmed Experimentally :: 0
            Transformed Into         :: unspecified
            Sequence Document Type   :: UNA
            SnapGene UUID            :: da1869e7-1e8d-475a-a050-d24a7c2a80c5
            Use Custom Sequence Name :: 1
            Sequence Type            :: Synthetic
            ##SnapGene_Notes-END##
            ##Imported_From-START##
            Path     :: C:\Users\TheGoldenLab\Downloads
            Filename :: addgene-plasmid-105715-sequence-207274.dna
            ##Imported_From-END##
FEATURES             Location/Qualifiers
     repeat_region   1..130
                     /note="<html><body>Functional equivalent of wild-type AAV2
                     ITR</body></html>"
                     /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="<html><body>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.</body></html>"
                     /bound_moiety="<html><body>Cre recombinase</body></html>"
                     /label="lox2272"
     protein_bind    712..745
                     /note="<html><body>Cre-mediated recombination occurs in
                     the 8-bp core sequence (GCATACAT).</body></html>"
                     /bound_moiety="<html><body>Cre recombinase</body></html>"
                     /label="loxP"
     CDS             complement(754..1473)
                     /created_by="ericszelenyi"
                     /label="AausFP1"
     Tag             complement(1474..1548)
                     /created_by="ericszelenyi"
                     /modified_by="ericszelenyi"
                     /label="ArgiNLS"
     misc_feature    complement(1549..1614)
                     /created_by="ericszelenyi"
                     /label="P2A peptide"
     CDS             complement(1615..3063)
                     /created_by="ericszelenyi"
                     /modified_by="ericszelenyi"
                     /label="hM4D(Gi)"
     Tag             complement(<3064..3168)
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /label="FLAG tag"
     protein_bind    complement(3181..3214)
                     /note="<html><body>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.</body></html>"
                     /bound_moiety="<html><body>Cre recombinase</body></html>"
                     /label="lox2272"
     protein_bind    complement(3265..3298)
                     /note="<html><body>Cre-mediated recombination occurs in
                     the 8-bp core sequence (GCATACAT).</body></html>"
                     /bound_moiety="<html><body>Cre recombinase</body></html>"
                     /label="loxP"
     misc_feature    3339..3583
                     /created_by="TheGoldenLab"
                     /modified_by="TheGoldenLab"
                     /label="WPRE3"
     primer_bind     complement(3395..3415)
                     /Tm=55
                     /Description="WPRE, reverse primer"
                     /label="WPRE-R"
     terminator      3583..3704
                     /created_by="TheGoldenLab"
                     /label="SV40 pA"
     misc_feature    3700..3705
                     /created_by="TheGoldenLab"
                     /label="HpaI"
     misc_feature    3711..3716
                     /created_by="TheGoldenLab"
                     /label="MfeI"
     repeat_region   3797..3926
                     /label="AAV2 ITR"
     rep_origin      4012..4467
                     /direction="RIGHT"
                     /note="<html><body>f1 bacteriophage origin of replication;
                     arrow indicates direction of (+) strand
                     synthesis</body></html>"
                     /label="f1 ori"
     primer_bind     complement(4099..4118)
                     /Tm=57
                     /Description="F1 origin, reverse primer"
                     /label="F1ori-R"
     primer_bind     4309..4330
                     /Tm=57
                     /Description="F1 origin, forward primer"
                     /label="F1ori-F"
     primer_bind     complement(4484..4503)
                     /Tm=54
                     /Description="pRS vectors, use to sequence yeast
                     selectable marker"
                     /label="pRS-marker"
     primer_bind     4603..4625
                     /Tm=66
                     /Description="pGEX vectors, reverse primer"
                     /label="pGEX 3'"
     primer_bind     complement(4663..4681)
                     /Tm=57
                     /Description="pBR322 vectors, upsteam of EcoRI site,
                     forward primer"
                     /label="pBRforEco"
     promoter        4749..4853
                     /gene="<html><body><i>bla</i></body></html>"
                     /label="AmpR promoter"
     Promoter        4784..4812
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="amp prom"
                     /Transferred_Similarity="100.00%"
                     /label="amp prom"
     CDS             join(4854..4922,4923..5714)
                     /gene="<html><body><i>bla</i></body></html>"
                     /note="<html><body>confers resistance to ampicillin,
                     carbenicillin, and related antibiotics</body></html>"
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVG
                     YIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLV
                     EYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTR
                     LDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGP
                     LLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQI
                     AEIGASLIKHW*"
                     /transl_table=1
                     /product="<html><body>β-lactamase</body></html>"
                     /label="AmpR"
     primer_bind     complement(5072..5091)
                     /Tm=58
                     /Description="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     rep_origin      5885..6472
                     /direction="RIGHT"
                     /note="<html><body>high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of replication</body></html>"
                     /label="ori"
     primer_bind     6373..6392
                     /Tm=56
                     /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 gccttaggcg taggcggtct tgcagtcgat
      781 ggctctgaac acttccacca cgttcatgtg gtcccgggtt tctttagggt ccttgctcag
      841 gatggtgtgg gcctgaatgt ggtgatagtg agggatgtcc acggggccac ctccgagagg
      901 ggtattcagc tgctggtgag cagcaatgat ctggcctccg ccgatcacgt cgtgcacaat
      961 gttgatgtag cacttgatgc cgttgttctg cacgtcgggc agcacgtaga tgcagtgcgg
     1021 gttgtagttg aactcaagtt tcttgcccag gatgttgccg cctttcttga agccgctgcc
     1081 gttcagggtc actctgttgt acacggagcc cttctcgtag gtcacttcgg ctctggtctt
     1141 gtacacgccg tcgttctcga aggtgatggt tctctcttgc acgtagccct caggcatggc
     1201 gctcttgggg tagtccttga tgtcgttggg atacttggcg aagcactggg cgccgtatgt
     1261 cactgtggtc aggatgctgc tccaaggcac aggcagttcg ccggttgtgc agatgaagct
     1321 ggcctcgatc tttccggtat tggcgtcgcc gtgtcctttg cctctgacgc tgaacttcat
     1381 gccttccacg tcgccttcca tttccaccac ataggggatc ttctcgcgga acagcagggc
     1441 gccataagac atggtggcca caggggatcc cgttcgacgg cgccgccgcc ttctcacctt
     1501 ctttttcttt gctatgctca tctgcttctg gggagccacc cttttcatag gaccggggtt
     1561 ttcttccacg tctcctgctt gctttaacag agagaagttc gtggctccgg atccgaccgg
     1621 tggatcccgc ctggcagtgc cgatgttccg atactggcac agcagcaggt gccggaaggt
     1681 ctttttaaag gtggcgttgc acagagcata gcaggcaggg ttgatggtgc tgttgacgta
     1741 gcagagccag tagccaatgg accacaccgt gtcagggatg cagctctggc agaaggtgtt
     1801 caccaggacc atgacgttgt agggcgtcca ggtgaggatg aaggccagca gaatggcaaa
     1861 gatcgttcgt gtcactttgc gctcccgggc cgccatctgc cgcttcttgc gcacctggtt
     1921 gcgagcgatg ctggcgaact tgcgggccac gttggccgca gggcgcatgc cagccggcgt
     1981 ggcaggcaca atctcaatgg ctgtcacaca ctcattgcct gtctgcttcg tcacaatctg
     2041 gatcttggac catctggagg ctgggttgag ggcccgcggc tgcaggggag gggcgggcat
     2101 ggcgggcgtg gtggcctctg tggtggacag ctctgtggct gggcgttcct tggtgttctg
     2161 ggtggcactg cctgagctgg actcattgga agtgtcctta tcagccacgg ggcgcggtgg
     2221 cggtggcagc gctggcgggg gggcctcctc cagcttgcca ttgcgcagct cctcccgggc
     2281 ggcctccccg ggcgggggct tcttgacgct ctgcttcatt agtgggctct tgaggaaggc
     2341 cagcgtcttg gctttcttct ccttcgggcc ctcgggccgg tgcttgtgga ctcggctgcg
     2401 actggccagg gagatgtgga tgtacagcac cgtcatgatg accacaggca ggtagaagcc
     2461 agcaatggct gtgccaaagg tcactgctgg gttggacagg aactggatga agcactggtt
     2521 gtcgggcacc gtccgcttac ccaccacaaa ctgccagaac aagatggcag gcgcccagag
     2581 cacgaaggac agtacccagg cagcagcaat catgaggcct gccatcttgg tggtgcgccg
     2641 ggcagggtag gtgagaggct tggtgacgca gaagtagcgg tcaaagctga tgatgagaag
     2701 gttcatgacg gaggcgttgc tcaccacgca gtccagggcc agccacaggt cgcagaccac
     2761 ggcgcccagg ggccagtagc ccttgatgat gtacacggtg tagaggttca tggagaaggc
     2821 gcctatgatg agatcagcac acgccaggct gaagaggaag tagttgttga ctgtctgcag
     2881 ctgcctgttg accttgatgg acagcatcac caggatgttg cccacgacag tcaccaggct
     2941 cagggagcct gtcactgtgg caatgaagac catttccacc gtctcatagc gattgtggga
     3001 tgatgacgtg accaggcgca cggactgatt gcccgagctg ccattgacag gtgtgaagtt
     3061 ggcatttaaa tcttcttctg ataataattt ttgttctaat gcatctcgag tggatccgag
     3121 ctctgaattc tcaccgatat ctttatcatc atcatcttta taatccatgg tggcgctagc
     3181 ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg
     3241 ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga
     3301 attcgatatc aagcttatcg atatcgaatt ccgctcgaga taatcaacct ctggattaca
     3361 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3421 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3481 ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc tgccttgccc
     3541 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttt atttgtgaaa
     3601 tttgtgatgc tattgcttta tttgtaacca tctagcttta tttgtgaaat ttgtgatgct
     3661 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3721 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcgg ccgccacgtg
     3781 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3841 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3901 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3961 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4021 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4081 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4141 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4201 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4261 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4321 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     4381 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4441 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4501 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4561 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4621 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4681 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4741 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4801 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4861 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4921 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4981 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5041 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5101 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5161 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5221 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5281 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5341 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5401 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5461 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5521 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5581 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5641 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5701 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5761 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5821 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5881 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5941 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6001 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6061 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6121 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6181 ataaggcgca gcggtcgggc tgaacggggg ttcgtgcaca cagcccagct tggagcgaac
     6241 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6301 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6361 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6421 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6481 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
