LOCUS Exported 6645 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses synaptoPAC, a fusion of synaptophysin, mScarlet, and bPAC in neurons. ACCESSION . VERSION . KEYWORDS pAAV-syn-synaptoPAC-minWPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6645) TITLE SynaptoPAC, an Optogenetic Tool for Induction of Presynaptic Plasticity JOURNAL Unpublished REFERENCE 2 (bases 1 to 6645) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6645 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 71..211 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 71..200 /label=AAV2 ITR rep_origin 286..741 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(373..392) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 583..604 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(758..777) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 877..899 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(937..955) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1023..1127 /gene="bla" /label=AmpR promoter CDS 1128..1988 /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(1346..1365) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2159..2747 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2648..2667 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 2809..2938 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 2970..3417 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 4395..5090 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(4528..4546) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 4796..4815 /label=mCherry-F /note="mCherry, forward primer" CDS 6159..6188 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 6198..6215 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(6320..6340) /label=WPRE-R /note="WPRE, reverse primer" ORIGIN 1 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcac ggtgcggacc 61 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg 121 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca 181 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg 241 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg 301 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 361 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 421 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 481 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga 541 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 601 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga 661 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 721 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat 781 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc 841 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt 901 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat 961 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg 1021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 1081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 1141 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 1201 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 1261 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 1321 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 1381 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 1441 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 1501 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 1561 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 1621 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 1681 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 1741 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 1801 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 1861 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 1921 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 1981 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 2041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 2101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 2161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 2221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 2281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 2341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 2401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 2461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 2521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 2581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 2641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 2701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 2761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc 2821 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc 2881 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc 2941 ggccgcacgc gtagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg 3001 cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc 3061 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc 3121 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg 3181 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact 3241 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac 3301 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca 3361 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg 3421 ctagcgccac catgctgctg ctggcagaca tggacgtggt gaatcagctg gtggctgggg 3481 gtcagttccg ggtggtcaag gagccccttg gcttcgtgaa ggtgctgcag tgggtctttg 3541 ccatcttcgc ctttgctacg tgcggcagct acaccggaga gcttcggctg agcgtggagt 3601 gtgccaacaa gacggagagt gccctcaaca tcgaagtcga atttgagtac ccattcaggc 3661 tgcaccaagt gtactttgat gcaccctcct gcgttaaagg gggcactacc aagatcttcc 3721 tagttggtga ctactcctcc tcggctgaat tctttgtcac cgtggctgtg tttgccttcc 3781 tctactccat gggggccctg gccacctaca tcttcctgca gaacaagtac cgagagaaca 3841 acaaagggcc aatgatggac ttcctggcca cagcagtgtt cgctttcatg tggctagtta 3901 gctcatccgc ctgggccaaa ggcctgtccg atgtgaagat ggccactgac ccagagaaca 3961 ttatcaagga gatgcctatg tgccgccaga caggaaacac atgcaaggaa ctgagggacc 4021 ctgtgacttc aggactcaac acctcggtgg tgtttggctt cctgaacctg gtgctctggg 4081 ttggcaacct atggttcgtg ttcaaggaga caggctgggc cgccccattc atgcgcgcac 4141 ctccaggcgc cccagaaaag caaccagctc ctggcgatgc ctacggcgat gcgggctatg 4201 ggcagggccc cggaggctat gggccccagg actcctacgg gcctcagggt ggttatcaac 4261 ccgattacgg gcagccagcc agcggcggtg gcggtggcta cgggcctcag ggcgactatg 4321 ggcagcaagg ctacggccaa cagggtgcgc ccacctcctt ctccaatcag atggaagggt 4381 taactggctc caccatggtg agcaagggcg aggcagtgat caaggagttc atgcggttca 4441 aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg 4501 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc 4561 ccttctcctg ggacatcctg tcccctcagt tcatgtatgg ctccagggcc ttcaccaagc 4621 accccgccga catccccgac tactataagc agtccttccc cgagggcttc aagtgggagc 4681 gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc tccctggagg 4741 acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg 4801 taatgcagaa gaagacaatg ggctgggaag cgtccaccga gcggttgtac cccgaggacg 4861 gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg 4921 cggacttcaa gaccacctac aaggccaaga agcccgtgca gatgcccggc gcctacaacg 4981 tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg 5041 aacgctccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggatccatga 5101 tgaagcggct ggtgtacatc agcaagatca gcggccacct gagcctggaa gagatccagc 5161 ggatcggcaa ggtgtccatc aagaacaacc agcgggacaa catcaccggc gtgctgctgt 5221 acctgcaggg cctgttcttc cagatcctgg aaggcgagaa cgagaaggtg gacaagctgt 5281 acaagaaaat cctggtggac gaccggcaca ccaacatcct gtgcctgaaa accgagtacg 5341 acatcaccga ccggatgttc cccaactggg ccatgaaaac catcaacctg aacgagaaca 5401 gcgagctgat gatccagccc atcaagagcc tgctgcagac catcacccag agccaccggg 5461 tgctggaaaa gtacatgccc gccagagtga tctacctgat caaccagggc atcaaccccc 5521 tgaccgtgga gccccagctg gtcgagaaga tcatcttctt cagcgacatc ctggccttca 5581 gcaccctgac cgagaagctg cccgtgaacg aggtggtcat cctggtcaac cggtacttca 5641 gcatctgcac ccggatcatc agcgcctacg gcggcgaagt gaccaagttc atcggcgact 5701 gcgtgatggc cagcttcacc aaagagcagg gcgacgccgc catccggacc agcctggaca 5761 tcatctccga gctgaagcag ctgcggcacc acgtggaggc caccaacccc ctgcacctgc 5821 tgtacaccgg catcggcctg agctacggcc acgtgatcga gggcaacatg ggcagcagcc 5881 tgaagatgga ccacaccctg ctgggggacg ccgtgaacgt ggccgccagg ctggaagccc 5941 tgacaagaca gctgccctac gccctggcct ttacagccgg cgtgaagaag tgctgccagg 6001 cccagtggac cttcatcaac ctgggagccc accaggtcaa gggcaagcag gaagccatcg 6061 aggtctacac cgtgaatgag gcccagaagt actacgacac cctgcagatc acccagctga 6121 tccggcagac cctggaaaac gacaagccgc ggacgtacga gcaaaagctg atctcggagg 6181 aggacctgaa gggatcgcac caccatcacc accatgcata accttcgaat taattaaggc 6241 cgcgactcta gaattccgct cgagataatc aacctctgga ttacaaaatt tgtgaaagat 6301 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc 6361 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct 6421 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 6481 ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg 6541 ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc tttatttgta 6601 accattataa gctgcaataa acaagttaac aacaacaatt gcatt //