LOCUS Exported 6255 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Axon-targeted yellow protein calcium sensor expressed under human Synapsin1 promoter, Cre mediated flip-excision switch, for AAV production or direct transfection, slow variant. ACCESSION . VERSION . KEYWORDS pAAV-syn-FLEX-axon-jYCaMP1s SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6255) AUTHORS Mohr MA, Bushey D, Aggarwal A, Marvin JS, Kim JJ, Marquez EJ, Liang Y, Patel R, Macklin JJ, Lee CY, Tsang A, Tsegaye G, Ahrens AM, Chen JL, Kim DS, Wong AM, Looger LL, Schreiter ER, Podgorski K TITLE jYCaMP: an optimized calcium indicator for two-photon imaging at fiber laser wavelengths. JOURNAL Nat Methods. 2020 May 25. pii: 10.1038/s41592-020-0835-7. doi: 10.1038/s41592-020-0835-7. PUBMED 32451475 REFERENCE 2 (bases 1 to 6255) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6255 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 39..168 /label=AAV2 ITR intron 761..893 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 876..895 /label=pMT2-F /note="Synthetic intron, forward primer" protein_bind complement(906..939) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(1002..1035) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind 1612..1631 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1871..1892 /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1998..2019) /label=EGFP-C /note="EGFP, forward primer" CDS complement(2232..2288) /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRVIGRLSS" regulatory 2300..2309 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(2316..2339) /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS complement(2343..2375) /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS complement(2379..2396) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" protein_bind 2493..2526 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 2597..2630 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2702..3290 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2755..2775) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3173..3184) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3368..3489 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3379..3398) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3433..3452 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3846..3863) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4017..4605) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4097..4116) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4776..5636) /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 5399..5418 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5637..5741) /gene="bla" /label=AmpR promoter rep_origin complement(5767..6222) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5904..5925) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6116..6135 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg 61 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg 121 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga 181 ttaacccgcc atgctactta tctacgtagc catgctctag gaagatccga gctccagtgt 241 gctggaattc gcccttttca gtatttaaat 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 gagcgcagct gtgctcctgg gcaccgcgca gtccgccccc gcggctcctg gccagaccac 721 ccctaggacc ccctgcccca agtcgcagcc gttggatcag gtaagtatca aggttacaag 781 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt 841 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgatga 901 tccccataac ttcgtataaa gtatcctata cgaagttata tcaaaatagg aagaccaatg 961 cttcaccatc gacccgaatt gccaagcatc accatcgaca cataacttcg tataatgtat 1021 gctatacgaa gttatactag tgctagcgga tcctacttcg ctgtcatcat ttgtacaaac 1081 tcttcgtagt ttacctgacc atccccatcg atgtctgctt ccctgatcat ttcatcaacc 1141 tcttcatctg ttaacttctc tccaaggttt gtcatcacgt ggcgaagctc tgctgcactg 1201 atgtagccat tgccatcctt atcaaacaca ccgaacgctt ctctaatttc ttcttccgtg 1261 tccctgtatt tcatttttct tgccatcatt gtcaggaact cagggaagtc gattgtgccg 1321 tcaccgtcgg catctacttc attgatcatg tcctgcagct ctgcttctgt ggggttctgc 1381 cccagagacc gcatcaccgt ccccagctcc ttggttgtta ttgtcccatc cccgtccttg 1441 tcaaataggg agaaaagctc tttaaattct gcgatctgct cttcagtcag ttggtcagga 1501 aggttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgccctta 1561 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag 1621 ttgccgtcgt ccttgaagaa gatggtgcgc tcctgcacgt agccttcggg catggcggac 1681 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg caggccgtag 1741 cccagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcaaatcagc 1801 ttcagggtca gcttgccgta ggtggcatcg ccctcaccct cgccggacac gctgaacttg 1861 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc 1921 tcgcccttgc tcaccatgct ccctccggta ccgcccttgt acagctcatc catgcccaga 1981 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg 2041 tctttgctaa gcacgggctg gtagctcagg tagtggttgt cgggcagcag cacggggccg 2101 tcgccgatgg gggtgttctg ctggtagtga taggcgagct gcacgccgcc gtcctcgatg 2161 ttgtggcgga tgtggaagtt cgccttgatg ccgttcttct gcttgtcggc ggtgatatag 2221 acgttctcga gtgagctcag ccgacctatc actctgactg cgtgacctgt cttattccac 2281 ttacgacgtg atgagtcgac catggtggcg agatccttat cgtcatcgtc gtacagatcc 2341 cgacccattt gctgtccacc agtcatgcta gccataccgt gatgatgatg gtgatgagaa 2401 ccaatctttt ggtcctcatc attcttttca acctgtttgg ttcttctcat acagcacagc 2461 atggtggcga tatcaccggt ggtaccacta gtataacttc gtataggata ctttatacga 2521 agttatcatt gggattcttc ctattttgat ccaagcatca ccatcgaccc tctagtccaa 2581 atctcaccat cgacccataa cttcgtatag catacattat acgaagttat gtccctcgaa 2641 gaggttcgaa acactgaaaa gggccaattc tgcaaatgat ccactcgaga agcttatcga 2701 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 2761 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 2821 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 2881 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 2941 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 3001 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 3061 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 3121 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 3181 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 3241 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg 3301 tcgacccggg cggccgctcg agcatgcatc tagagtcgac ccgggcggcc gcttcgagca 3361 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa 3421 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat 3481 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg 3541 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct 3601 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct 3661 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg acgcccgggc 3721 tttgcccggg cggcgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag 3781 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 3841 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 3901 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 3961 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 4021 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 4081 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 4141 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 4201 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 4261 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 4321 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 4381 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 4441 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 4501 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 4561 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 4621 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 4681 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 4741 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 4801 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 4861 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 4921 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 4981 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 5041 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 5101 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 5161 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 5221 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 5281 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 5341 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 5401 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 5461 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 5521 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 5581 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 5641 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 5701 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 5761 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 5821 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 5881 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 5941 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 6001 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 6061 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 6121 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 6181 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc 6241 tgcgcaactg ttggg //