LOCUS Exported 6863 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Synaptic Tag for Mapping Activity. ACCESSION . VERSION . KEYWORDS SynTagMA_pre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6863) AUTHORS Perez-Alvarez A, Fearey BC, O'Toole RJ, Yang W, Arganda-Carreras I, Lamothe-Molina PJ, Moeyaert B, Mohr MA, Panzera LC, Schulze C, Schreiter ER, Wiegert JS, Gee CE, Hoppa MB, Oertner TG TITLE Freeze-frame imaging of synaptic activity using SynTagMA. JOURNAL Nat Commun. 2020 May 18;11(1):2464. doi: 10.1038/s41467-020-16315-4. PUBMED 32424147 REFERENCE 2 (bases 1 to 6863) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6863 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 47..176 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 221..668 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 1732..1749 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 1756..2196 /codon_start=1 /product="calcium-binding protein" /label=calmodulin /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK" CDS 2917..2973 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" misc_feature 3036..3624 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3089..3109) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3507..3518) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3656..4132 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3855..3874) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4172..4312 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4172..4301 /label=AAV2 ITR rep_origin 4387..4842 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4474..4493) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4684..4705 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4859..4878) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4978..5000 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5038..5056) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5124..5228 /gene="bla" /label=AmpR promoter CDS 5229..6089 /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(5447..5466) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6260..6848 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6749..6768 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg 61 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc 121 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg 181 ccgcacgcgt gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg ggttttagga 241 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc 301 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga 361 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc 421 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc 481 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg 541 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg 601 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga 661 gagcgcagtc gagaaggtac cggatccccg ggtaccggta tggacgtggt gaatcagctg 721 gtggctgggg gtcagttccg ggtggtcaag gagccccttg gcttcgtgaa ggtgctgcag 781 tgggtctttg ccatcttcgc ctttgctacg tgcggcagct acaccggaga gcttcggctg 841 agcgtggagt gtgccaacaa gacggagagt gccctcaaca tcgaagtcga atttgagtac 901 ccattcaggc tgcaccaagt gtactttgat gcaccctcct gcgttaaagg gggcactacc 961 aagatcttcc tagttggtga ctactcctcc tcggctgaat tctttgtcac cgtggctgtg 1021 tttgccttcc tctactccat gggggccctg gccacctaca tcttcctgca gaacaagtac 1081 cgagagaaca acaaagggcc aatgatggac ttcctggcca cagcagtgtt cgctttcatg 1141 tggctagtta gctcatccgc ctgggccaaa ggcctgtccg atgtgaagat ggccactgac 1201 ccagagaaca ttatcaagga gatgcctatg tgccgccaga caggaaacac atgcaaggaa 1261 ctgagggacc ctgtgacttc aggactcaac acctcggtgg tgtttggctt cctgaacctg 1321 gtgctctggg ttggcaacct atggttcgtg ttcaaggaga caggctgggc cgccccattc 1381 atgcgcgcac ctccaggcgc cccagaaaag caaccagctc ctggcgatgc ctacggcgat 1441 gcgggctatg ggcagggccc cggaggctat gggccccagg actcctacgg gcctcagggt 1501 ggttatcaac ccgattacgg gcagccagcc agcggcggtg gcggtggcta cgggcctcag 1561 ggcgactatg ggcagcaagg ctacggccaa cagggtgcgc ccacctcctt ctccaatcag 1621 atgaagggcg tggagcccaa gacctactgc tactacagca gcaagcttat gctgcagaac 1681 gagcttgctc ttaagttggc tggacttgat attaacaaga ctggaggttc tcatcatcac 1741 caccaccatg gatccgacca actgactgaa gagcagatcg cagaatttaa agaggctttc 1801 tccctatttg acaaggacgg ggatgggaca ataacaacca aggagctggg gacggtgatg 1861 cggtctctgg ggcagaaccc cacagaagca gagctgcagg acatgatcaa tgaagtagat 1921 gccgacggtg acggcacaat cgacttccct gagttcctga caatgatggc aagaaaaatg 1981 aaagacacag acagtgaaga agaaattaga gaagcgttcc gtgtgtttga taaggatggc 2041 aatggctaca tcagtgcagc agagcttcgc cacgtgatga caaaccttgg agagaagtta 2101 acagatgaag aggttgatga aatgatcagg gaagcagaca tcgatgggga tggtcaggta 2161 aactacgaag agtttgtagt gatgatgaca gcgaagctcg agtgcgagaa aatctatgtg 2221 cgtgatggag tgctgacggg tgatattcat atgatcttgt tgcttgaagg aaatgcccat 2281 taccgatgtg acttcagaac tacttacaaa gctaaggaga agggtgtcaa gttaccaggc 2341 gtgcactatg tggaccacac cattgagatt ttaagccatg acaaagatta caacaaggtt 2401 aagatctatg agtatgctgt tgctcattct ggattgcctg acaatgccag acgaggcggt 2461 accggcggat ccatggtgag tgcgattaag ccagacatga agatcaaact gcgtatggaa 2521 ggcaacgtaa acgggcacca ctttgtgatc gacggagatg gtacaggcaa gccttatgag 2581 ggaaaacaga ccatggatct tgaagtcaaa gagggcggac ctctgccttt tgcctttgat 2641 atcctgacca ctgcattcca ttacggcaac agggtattcg tgaaatatcc agacaacata 2701 caagactatt ttaagcagtc gtttcctaag gggtattcgt gggaacgaag catgactttc 2761 gaagacgggg gcatttgcta tgccagaagc gacataacaa tggaagggga cactttctat 2821 aataaagttc gattttatgg taccaacttt cccgccaatg gtccagttat gcagaagaag 2881 acgctgaaat ggatgccgag ctggacgcgt tcatcacgtc gtaagtggaa taagacaggt 2941 cacgcagtca gagctatagg tcggctgagc tcataatgta caagttctgc tacgagaacg 3001 aggtgtaatg agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt 3061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 3121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 3181 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 3241 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 3301 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 3361 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 3421 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc 3481 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 3541 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 3601 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg 3661 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc 3721 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta 3781 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg 3841 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac 3901 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg 3961 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt 4021 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc 4081 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt 4141 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact 4201 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg 4261 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg 4321 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac 4381 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 4441 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc 4501 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 4561 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta 4621 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 4681 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg 4741 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa 4801 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta 4861 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg 4921 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg 4981 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc 5041 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag 5101 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 5161 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 5221 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 5281 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 5341 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 5401 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 5461 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 5521 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 5581 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 5641 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 5701 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 5761 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 5821 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 5881 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 5941 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 6001 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 6061 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 6121 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 6181 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 6241 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 6301 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 6361 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 6421 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 6481 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 6541 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 6601 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 6661 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 6721 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 6781 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 6841 tatggaaaaa cgccagcaac gcg //