LOCUS Exported 5929 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION soluble GCaMP expression . ACCESSION . VERSION . KEYWORDS pyc128m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5929) AUTHORS Chen Y, Jang H, Spratt PWE, Kosar S, Taylor DE, Essner RA, Bai L, Leib DE, Kuo TW, Lin YC, Patel M, Subkhangulova A, Kato S, Feinberg EH, Bender KJ, Knight ZA, Garrison JL TITLE Soma-Targeted Imaging of Neural Circuits by Ribosome Tethering. JOURNAL Neuron. 2020 Jun 5. pii: S0896-6273(20)30351-2. doi: 10.1016/j.neuron.2020.05.005. PUBMED 32574560 REFERENCE 2 (bases 1 to 5929) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5929 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..448 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 475..484 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 481..1833 /codon_start=1 /product="improved fluorescent protein-based calcium sensor (Chen et al., 2013)" /label=GCaMP6m /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK" regulatory 577..586 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind 867..888 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(994..1015) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1255..1274) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 1874..2462 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1927..1947) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2345..2356) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2494..2970 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2693..2712) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3010..3150 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3010..3139 /label=AAV2 ITR rep_origin 3225..3680 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3312..3331) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3522..3543 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3697..3716) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3816..3838 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3876..3894) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3962..4066 /gene="bla" /label=AmpR promoter CDS 4067..4927 /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(4285..4304) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5098..5686 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5587..5606 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 5748..5877 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 61 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 121 gagggggagg ggaaacagga tgcggcgagg ctcgtgcgca 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 ctagaggatc cggcgccacc 481 atgggttctc atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg 541 ggtcgggatc tgtacgacga tgacgataag gatctcgcca ccatggtcga ctcatcacgt 601 cgtaagtgga ataagacagg tcacgcagtc agagctatag gtcggctgag ctcactcgag 661 aacgtctata tcaaggccga caagcagaag aacggcatca aggcgaactt caagatccgc 721 cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc 781 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc caaactttcg 841 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 901 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag 961 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 1021 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc 1081 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 1141 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 1201 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac 1261 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 1321 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 1381 aacctgccgg accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta 1441 tttgacaagg acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct 1501 ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac 1561 ggtgacggca caatcgactt ccctgagttc ctgacaatga tggcaagaaa agggagctac 1621 agggacacgg aagaagaaat tagagaagcg ttcggtgtgt ttgataagga tggcaatggc 1681 tacatcagtg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat 1741 gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac 1801 gaagagtttg tacaaatgat gacagcgaag tgaggcgcgc ccctgcaggg aattcgatat 1861 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct 1921 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc 1981 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct 2041 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga 2101 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc 2161 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac 2221 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt 2281 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt 2341 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc 2401 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc 2461 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca 2521 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa 2581 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg 2641 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg 2701 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt 2761 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg 2821 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc 2881 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag 2941 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg 3001 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc 3061 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag 3121 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc 3181 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg 3241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 3301 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 3361 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 3421 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac 3481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 3541 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat 3601 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa 3661 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata 3721 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct 3781 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt 3841 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata 3901 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt 3961 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 4021 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 4081 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 4141 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 4201 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 4261 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 4321 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 4381 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 4441 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 4501 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 4561 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 4621 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 4681 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 4741 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc 4801 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 4861 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 4921 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 4981 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 5041 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 5101 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 5161 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 5221 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 5281 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 5341 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 5401 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 5461 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 5521 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 5581 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga 5641 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 5701 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct 5761 gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct 5821 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg 5881 gccgcacgcg tttaattaag tgtctagact gcagagggcc ctgcgtatg //