LOCUS Exported 7252 bp ds-DNA circular SYN 20-JAN-2020 DEFINITION . ACCESSION . VERSION . KEYWORDS pAAV_hsyn_synzip_GFP no GCaMP6f SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 7252) AUTHORS Trial User TITLE Direct Submission JOURNAL Exported Monday, Jan 20, 2020 from SnapGene 5.0.7 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7252 /organism="recombinant plasmid" /mol_type="other DNA" promoter 1..448 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" /note="color: #ffffff; direction: RIGHT" regulatory 463..472 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /note="color: #a6acb3" CDS 469..1185 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /note="This feature has 3 segments: 1: 469 .. 471 / #05fd14 2: 472 .. 474 / #05fd14 / 1a 3: 475 .. 1185 / #05fd14" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" misc_feature 1225..1746 /label=SYNAPTOZIP /note="color: #84a4c0; direction: RIGHT" misc_feature 1624..1713 /label=ZIP /note="color: #ff00ff; direction: RIGHT" CDS 1678..1689 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /note="color: #cc99b2 Cleavage site after base 1689" /translation="WELQ" CDS 1714..1743 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /note="color: #cc99b2" /translation="EQKLISEEDL" misc_feature 1747..3172 /label=3'UTR VAMP2 /note="color: #339966" misc_feature 3181..3687 /label=insert from pLenti_mCherry_SynaptoZip.xdna /note="note=""Fragment extracted from pLenti_mCherry_SynaptoZip.xdna [3211 nt] : (AgeI[5145] / SacII[8359])" /note="color: #84a4c0; direction: RIGHT" misc_feature 3186..3774 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" /note="color: #ffffff" CDS complement(3657..3668) /label=FactorXa site /note="color: #008000; direction: LEFT" polyA_signal 3805..4285 /label=hGH polyA signal /note="color: #ffc0cb; direction: RIGHT" repeat_region 4322..4462 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" /note="color: #ffe4c4" rep_origin 4537..4992 /direction=RIGHT /label=M13 origin /note="color: #4682b4" promoter 5309..5337 /label=Amp prom /note="color: #4682b4; direction: RIGHT" CDS 5577..6236 /label=AmpR /note="color: #8fbc8f; direction: RIGHT" rep_origin 6388..7016 /direction=RIGHT /label=ColE1 origin /note="color: #4682b4" repeat_region 7060..7200 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" /note="color: #ffe4c4" ORIGIN 1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 121 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 181 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 241 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 301 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 361 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg 421 aggagtcgtg tcgtgcctga gagcgcagtc gagaaaccgg tcgccaccat ggtgagcaag 481 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 541 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc 601 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 661 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 721 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac 781 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 841 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 901 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg 961 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag 1021 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc 1081 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc 1141 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg ccggactcag 1201 atctcgagct caagcttcga attctcggct accgctgcca ccgtcccgcc tgccgccccg 1261 gccggcgagg gtggcccccc tgcacctcct ccaaatctta ccagtaacag gagactgcag 1321 cagacccagg cccaggtgga tgaggtggtg gacatcatga gggtgaatgt ggacaaggtc 1381 ctggagcgag accagaagct atcggaactg gatgatcgcg cagatgccct ccaggcaggg 1441 gcctcccagt ttgaaacaag tgcagccaag ctcaagcgca aatactggtg gaaaaacctc 1501 aagatgatga tcatcttggg agtgatttgc gccatcatcc tcatcatcat catcgtttac 1561 ttcagcactt ctgtccctga gggtaagggc gtggagccaa agacctactg ctactacagc 1621 agcgctcagc tggagaagga gctgcaggcc ctggagaagg agaacgcaca actagaatgg 1681 gaacttcaag cgctcgaaaa agaactggct caggaacaaa aactgatcag cgaggaggac 1741 ctgtgacctg aggttcccat cacctctctc tctcaccctc tgcccccttt cctctttcct 1801 gttcagcccc aagtcccttc atttgcatct gctatgcaat agcccctctc ctccttccct 1861 tggatttaac caatccttcc cctatcttcc ctgtacaatt ccagaccctc cccaaacaga 1921 gcaaacccca cagaaacaaa caaaacaccc cacctgtcta ggcttccata ggttgcatct 1981 ttgtatccct tgggagcctc taagactggt cctacttggt cctaagaatc ccaaggtttt 2041 ctgggagctt ccaacatgtt gattagcata tcatttgcat acactgtctt ttttttttcc 2101 ctgggtttct tcctttctgt tccattactc ttcactcttc tgtgtttttt ttttttcttt 2161 ctttcttttc ggaagagtta gtttccattg gtcctcttat cacactttta tttgcacgac 2221 attatctgca ggtggtgggg agcctgggct tttggggaac taggacccca aagttgcctc 2281 ctcccagccc ctctagtcta gtttgcttcc cttaccccat tttccacacc tcttgtaccc 2341 tcctctccct cccccagctg gtgtgtaagt gtcttggagt tcagtgtgtt atgatggacc 2401 aataattctg ccacttgggg tctctcccta cattcctgct ccccagtttt cgtgtggggc 2461 attcagctga cattcccggg ggtctccctc cctcccatct gcctgatctg ctgcctcctc 2521 ccaccaggag aactggaggc tgaccacaat ctggttcttt gaggaggggt ggctctagtg 2581 tgtgtggggg tcatcactgc cttggggagg agtgggacaa aacagaaccc cccctaattc 2641 ctgcctgaaa tctctggcct catccttgct agaggttgga ctgaaaactt tcctccccaa 2701 tcctgggggg gggtattacc ccccatcact gcccagctcc tctgactgcc cccctgtatt 2761 cagggtgggg gtactagtca ctgccaatat gtgtatggga cttgctggag gatggggatc 2821 cttgtccttc tctagggctg ctgagccctg agagagagag agagagagag agagagagag 2881 agagagagag agagagagag agggaagcaa gctatttccc tgttgggtac aatgatagag 2941 aaaggatcta atctttaaaa atggcacagt tttaggggtc tgagggtaca gccccttaac 3001 ctgcctcttg gggggggtgt tccccaaact cttcccccac acacaccagg ttttctgtgt 3061 ggaggggaac caaggagatc taaactgtgg tgtgaaaggg taggagagat gctgggggtg 3121 ggggtgcttg tgttttagac ccccaatatt atcccagtgt cccctgcctg aattctgcag 3181 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 3241 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 3301 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 3361 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 3421 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 3481 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 3541 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 3601 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 3661 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 3721 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat 3781 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct 3841 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg 3901 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg 3961 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag 4021 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat 4081 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta 4141 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc 4201 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac 4261 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg 4321 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag 4381 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag 4441 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc 4501 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa 4561 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 4621 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 4681 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 4741 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 4801 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 4861 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct 4921 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 4981 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 5041 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 5101 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 5161 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg 5221 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa 5281 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 5341 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 5401 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 5461 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 5521 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 5581 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 5641 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 5701 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 5761 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 5821 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 5881 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 5941 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 6001 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 6061 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 6121 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 6181 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 6241 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 6301 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 6361 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 6421 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 6481 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 6541 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 6601 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 6661 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 6721 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 6781 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 6841 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 6901 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 6961 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 7021 tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc 7081 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg 7141 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct 7201 gcggccgcac gcgtttaatt aagtgtctag actgcagagg gccctgcgta tg //