LOCUS       pAAV-Ef1a-ArgiNLS-mkate2        5426 bp    DNA     circular UNA 29-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...2pr-ek0cwe8
VERSION     urn.local...2pr-ek0cwe8
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     ##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        231..1409
                     /note="<html><body>strong constitutive promoter for human
                     elongation factor EF-1α</body></html>"
                     /label="EF-1α promoter"
     primer_bind     1357..1377
                     /Tm=57
                     /Description="Human elongation factor-1a promoter, forward
                     primer"
                     /label="EF1a-F"
     primer_bind     1411..1430
                     /%GC="55.0"
                     /Hairpin_Tm="52.3"
                     /Self_Dimer_Tm="9.5"
                     /Tm="57.4"
                     /Sequence="GTACCGTACCGGATCCTCTA"
                     /created_by="primer3"
                     /Transferred_From="GL_5"
                     /Transferred_Similarity="100.00%"
                     /label="GL_5"
     misc_feature    1433^1434
                     /Original_Bases="GGTACCG"
                     /label="GGTACCG"
                     /note="Geneious type: Editing History Deletion"
     sig_peptide     1442..1513
                     /created_by="TheGoldenLab"
                     /modified_by="ericszelenyi"
                     /label="ArgiNLS"
     CDS             1514..2209
                     /product="<html><body>monomeric far-red fluorescent
                     protein (<a
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/19143658""
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/19143658"">Shch
                     erbo et al., 2009</a>)</body></html>"
                     /transl_table=1
                     /codon_start=1
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVL
                     TATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADM
                     ALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHR*"
                     /note="<html><body>mammalian
                     codon-optimized</body></html>"
                     /label="mKate2"
     unsure          2213..2218
                     /created_by="TheGoldenLab"
                     /modified_by="TheGoldenLab"
                     /label="EcoRV-2"
     primer_bind     complement(2227..2249)
                     /%GC="43.5"
                     /Hairpin_Tm="33.2"
                     /Self_Dimer_Tm="5.9"
                     /Tm="57.3"
                     /Sequence="ATCCAGAGGTTGATTATCTCGAG"
                     /created_by="primer3"
                     /Transferred_From="GL_23"
                     /Transferred_Similarity="100.00%"
                     /label="GL_23"
     misc_feature    2232..2476
                     /created_by="TheGoldenLab"
                     /modified_by="TheGoldenLab"
                     /label="WPRE3"
     primer_bind     complement(2288..2308)
                     /Tm=55
                     /Description="WPRE, reverse primer"
                     /label="WPRE-R"
     terminator      2476..2597
                     /created_by="TheGoldenLab"
                     /label="SV40 pA"
     misc_feature    2593..2598
                     /created_by="TheGoldenLab"
                     /label="HpaI"
     misc_feature    2604..2609
                     /created_by="TheGoldenLab"
                     /label="MfeI"
     repeat_region   2690..2819
                     /label="AAV2 ITR"
     rep_origin      2905..3360
                     /direction="RIGHT"
                     /note="<html><body>f1 bacteriophage origin of replication;
                     arrow indicates direction of (+) strand
                     synthesis</body></html>"
                     /label="f1 ori"
     primer_bind     complement(2992..3011)
                     /Tm=57
                     /Description="F1 origin, reverse primer"
                     /label="F1ori-R"
     primer_bind     3202..3223
                     /Tm=57
                     /Description="F1 origin, forward primer"
                     /label="F1ori-F"
     primer_bind     complement(3377..3396)
                     /Tm=54
                     /Description="pRS vectors, use to sequence yeast
                     selectable marker"
                     /label="pRS-marker"
     primer_bind     3496..3518
                     /Tm=66
                     /Description="pGEX vectors, reverse primer"
                     /label="pGEX 3'"
     primer_bind     complement(3556..3574)
                     /Tm=57
                     /Description="pBR322 vectors, upsteam of EcoRI site,
                     forward primer"
                     /label="pBRforEco"
     promoter        3642..3746
                     /gene="<html><body><i>bla</i></body></html>"
                     /label="AmpR promoter"
     Promoter        3677..3705
                     /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(3747..3815,3816..4607)
                     /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(3965..3984)
                     /Tm=58
                     /Description="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     rep_origin      4778..5365
                     /direction="RIGHT"
                     /note="<html><body>high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of replication</body></html>"
                     /label="ori"
     primer_bind     5266..5285
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccgtacc ggatcctcta gagccaccat
     1441 gaaaagggtg gctccccaga agcagatgag catagcaaag aaaaagaagg tgagaaggcg
     1501 gcggcgccgt cgagtgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     1561 gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga
     1621 gggcacccag accatgagaa tcaaggcggt cgagggcggc cctctcccct tcgccttcga
     1681 catcctggct accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat
     1741 ccccgacttc tttaagcagt ccttccccga gggcttcaca tgggagagag tcaccacata
     1801 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     1861 ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa
     1921 aacactcggc tgggaggcct ccaccgagac cctgtacccc gctgacggcg gcctggaagg
     1981 cagagccgac atggccctga agctcgtggg cgggggccac ctgatctgca acttgaagac
     2041 cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga
     2101 cagaagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt
     2161 ggctgtggcc agatactgcg acctccctag caaactgggg cacagatgat aggatatcga
     2221 attccgctcg agataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2281 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2341 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg
     2401 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2461 gcactgacaa ttccgtggtg tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2521 ccatctagct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2581 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2641 atgtgggagg ttttttaaag cggccgccac gtgcggaccg agcggccgca ggaaccccta
     2701 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2761 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     2821 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2881 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2941 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     3001 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3061 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3121 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3181 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     3241 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     3301 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     3361 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     3421 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3481 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3541 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3601 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3661 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3721 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3781 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3841 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3901 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3961 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4021 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4081 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4141 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4201 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4261 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4321 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4381 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4441 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4501 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4561 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4621 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4681 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4741 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4801 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4861 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4921 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4981 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5041 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5101 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5161 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5221 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5281 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5341 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5401 cttttgctgg ccttttgctc acatgt
//
