LOCUS Exported 7138 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yellow fluorescent protein (YFP) – tagged rhodopsin adenylyl cyclase created from the rhodopsin guanylyl cyclase of Catenaria anguillulae with a neuron-specific promoter. Useful for raising intracellu. ACCESSION . VERSION . KEYWORDS pAAV hSyn1 YFP-CaRhAC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7138) AUTHORS Scheib U, Broser M, Constantin OM, Yang S, Gao S, Mukherjee S, Stehfest K, Nagel G, Gee CE, Hegemann P TITLE Rhodopsin-cyclases for photocontrol of cGMP/cAMP and 2.3 A structure of the adenylyl cyclase domain. JOURNAL Nat Commun. 2018 May 24;9(1):2046. doi: 10.1038/s41467-018-04428-w. PUBMED 29799525 REFERENCE 2 (bases 1 to 7138) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7138 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 423..552 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" rep_origin complement(614..1202) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(694..713) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1373..2233) /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 1996..2015 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2234..2338) /gene="bla" /label=AmpR promoter primer_bind 2406..2424 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2462..2484) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2584..2603 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(2620..3075) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2757..2778) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2969..2988 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 3150..3290 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3161..3290 /label=AAV2 ITR polyA_signal 3330..3806 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind 3588..3607 /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" misc_feature 3838..4426 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 3944..3955 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 4353..4373 /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6327..7043) /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(6369..6390) /label=EGFP-C /note="EGFP, forward primer" primer_bind 6718..6737 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6977..6998 /label=EGFP-N /note="EGFP, reverse primer" regulatory 7040..7049 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(7069..378) /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" ORIGIN 1 ccgcagcgca gatggtcgcg cccgtgcccc cctatctcgc gcctcgcgtg gtgcggtccg 61 gctgggccgg cggcggcgcg gacgcgacca aggtggccgg gaaggggagt ttgcggggga 121 ccggcgagtg acgtcagcgc gccttcagtg ctgaggcggc ggtggcgcgc gccgccaggc 181 gggggcgaag gcactgtccg cggtgctgaa gctggcagtg cgcacgcgcc tcgccgcatc 241 ctgtttcccc tccccctctc tgatagggga tgcgcaattt ggggaatggg ggttgggtgc 301 ttgtccagtg ggtcggggtc ggtcgtcagg taggcacccc caccccgcct catcctggtc 361 ctaaaaccca cttgcactca tacgcagggc cctctgcagt ctagacacac gcgtgcggcc 421 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga 481 ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca 541 gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 1021 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 1081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 1141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 1201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 1261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 1321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 1381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 1441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 1501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 1561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 1621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 1681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 1741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 1801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 1861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 1921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 1981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 2041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 2101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 2161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 2221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 2281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 2341 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 2401 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa 2461 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 2521 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact 2581 atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt 2641 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa 2701 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag 2761 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg 2821 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga 2881 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg 2941 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg 3001 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc 3061 cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg 3121 cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac 3181 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag 3241 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc 3301 ggtccgcacg tggttaccta caaaatcaga aggacaggga agggagcagt ggttcacgcc 3361 tgtaatccca gcaatttggg aggccaaggt gggtagatca cctgagatta ggagttggag 3421 accagcctgg ccaatatggt gaaaccccgt ctctaccaaa aaaacaaaaa ttagctgagc 3481 ctggtcatgc atgcctggaa tcccaacaac tcgggaggct gaggcaggag aatcgcttga 3541 acccaggagg cggagattgc agtgagccaa gattgtgcca ctgcactcca gcttggttcc 3601 caatagaccc cgcaggccct acaggttgtc ttcccaactt gccccttgct ccataccacc 3661 cccctccacc ccataatatt atagaaggac acctagtcag acaaaatgat gcaacttaat 3721 tttattagga caaggctggt gggcactgga gtggcaactt ccagggccag gagaggcact 3781 ggggaggggt cacagggatg ccacccgtag atctctcgag cagcgctcgg tatcgatgcg 3841 gggaggcggc ccaaagggag atccgactcg tctgagggcg aaggcgaaga cgcggaagag 3901 gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc tggattgagg gccgaaggga 3961 cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac ataggcgagc agccaaggaa 4021 aggacgatga tttccccgac aacaccacgg aattgtcagt gcccaacagc cgagcccctg 4081 tccagcagcg ggcaaggcag gcggcgatga gttccgccgt ggcaataggg agggggaaag 4141 cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt 4201 cagcaaacac agtgcacacc acgccacgtt gcctgacaac gggccacaac tcctcataaa 4261 gagacagcaa ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag 4321 catgatacaa aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa 4381 gaataccagt caatctttca caaattttgt aatccagagg ttgattatcg ataagcttat 4441 cactttcggg ctgtgaccca gtatgtttgc atctttccct ttcccttcac ttccatgaga 4501 tcgagtggtt gtgtcacgaa ctttcccttg atcatttggt atgtcgaatc cgagatgtgg 4561 atgtgtccag cctttgaagt cgattccatt cgcgaggcag tattcactgt atctcctacg 4621 aggtcccagt gtggattcaa atctccaaga actccggctg taactggtcc cgagttgagt 4681 ccgattcgga tgttgatcga ctctccagtg gctgtcttga atgtcttgat catctcgatg 4741 atatcgaggg cgaagttaac agctcgatca gcatgatctg gcacaacctc tggtgctcca 4801 gtaactccga ggtaagcatc tccgattgtc ttgaccttgt acactcccca tcgtttagcg 4861 atagcatcgt attcgagcca gagcttgttg agtgtggcca tcatatcttt tgtactagtt 4921 cgcgacgaga tcactgtgaa gttagtgata tccgagaaga acactgtgac tgactcgtac 4981 tctttggcct ctgtggcttg tccattcttg agttgttcga gaactccctc tggcatcatc 5041 ttttggagga tggcatcaca gtttgtgatt tgcgattcga gttcttcggc gatagctgtg 5101 atggcctcga ccttttgatt ttgggcttgc tcgactgtcg agttaacgag aacgagagtg 5161 aggaacacct tggctccgat atcgatcata gtgaatgagg cctctgtcat tgtgaacgag 5221 agcattcccg atgtgaaggc gaaccaactg agtgggaaga gtgtccaagt tgtcactgtc 5281 gagaatcgtg tccatcggag tccgttcact tcgatgttgc atggtgtttc tccgttgata 5341 gctccagtga acgatcgcca gagtgaatac accacgaacg agaagaaagc gcacgagaga 5401 atgttgaaga ggtttcccca tgggtatggt ggaagaacag ctccgaagaa tccgcacaca 5461 acaagagcgt aatctgagaa caccactccc caagcgttgt gatcagcttg tgtaatctcc 5521 gagatgagca ggatgagcac tgggcatgta gcaatccatt cgaggtatcg agctggctca 5581 actgggtttg agagtgttcc ctcgatagtt ggtgtgagtc ggagcatgat gaggatgtac 5641 gaactgaagg cgataatgtt cactccgagg aggcaaagcg aaagggcttt cttttctggg 5701 tttctagctc ctgcgaatcc catgtacgag aaggcgattc cgattccgaa tccaaaagct 5761 ccgtatccgt accaaatatc ggcccatcct cgattgtagg cttgtcgttc ttggatgaag 5821 agagcgtaga aagctccggc aactgtgaga atggcccaca tgatgaattg tcctgtgagg 5881 aacttgagtg atggtgttga gagcatcata tcccacgacc acatatcggc aatgtttgtt 5941 tggtattgtt ggagtgccga ttgttttgat ggcgatcctg gtcgcgaatt agctggcgac 6001 gaaacatttg acgacgaggc cgatgatgtc gatcgcacgt ttgacaccga gagcgagttg 6061 ttttgtcggt ttcgcatatt agcattcgac gaggctgtcg acgatcctgt cactttgatg 6121 tttggttgtg atggtgatgg aagaagtgaa gctcccgacg atctgaacga tccgtttcgt 6181 gtgagtggtt gctcaacgat tgggtgcttt gtgaccgagc acttgcaatc atcacagagt 6241 gtcgatgttg gcgagtagca atattctggg catgaacatc ctgagcaggc tccacggaga 6301 ttgttatcct tatccttctt ctcgagcttg tacagctcgt ccatgccgag agtgatcccg 6361 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc 6421 agggcggact ggtagctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg 6481 ggggtgttct gctggtagtg gtcggcgagc tgcacgccgc cgtcctcgat gttgtggcgg 6541 atcttgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgtgg 6601 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg 6661 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc 6721 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg 6781 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcagg 6841 ccgtagccca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag 6901 atcagcttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg 6961 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac 7021 agctcctcgc ccttgctcac catggtggcg gatccggtac cttctcgact gcgctctcag 7081 gcacgacacg actcctccgc tgcccaccgc agactgaggc agcgctgagt cgccggcg //