LOCUS Exported 7971 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION humanized, Neuron-specific promoter driven expression of the rhodopsin guanylyl cyclase from Catenaria anguillulae with a neuron-specific promoter. Useful for raising intracellular cAMP close to the m. ACCESSION . VERSION . KEYWORDS pAAV hSyn1 CaRhGC wt T2A tDimer SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7971) 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 7971) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7971 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 372..501 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" rep_origin complement(563..1151) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(643..662) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1322..2182) /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 1945..1964 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2183..2287) /gene="bla" /label=AmpR promoter primer_bind 2355..2373 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2411..2433) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2533..2552 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(2569..3024) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2706..2727) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2918..2937 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 3099..3239 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3110..3239 /label=AAV2 ITR polyA_signal 3279..3755 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind 3537..3556 /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind 3822..3838 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 3823..3839 /label=KS primer /note="common sequencing primer, one of multiple similar variants" misc_feature 3842..4430 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 3948..3959 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 4357..4377 /label=WPRE-R /note="WPRE, reverse primer" regulatory 5845..5854 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 5849..5858 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(5874..5927) /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" promoter complement(7847..323) /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" ORIGIN 1 gtccggctgg gccggcggcg gcgcggacgc gaccaaggtg gccgggaagg ggagtttgcg 61 ggggaccggc gagtgacgtc agcgcgcctt cagtgctgag gcggcggtgg cgcgcgccgc 121 caggcggggg cgaaggcact gtccgcggtg ctgaagctgg cagtgcgcac gcgcctcgcc 181 gcatcctgtt tcccctcccc ctctctgata ggggatgcgc aatttgggga atgggggttg 241 ggtgcttgtc cagtgggtcg gggtcggtcg tcaggtaggc acccccaccc cgcctcatcc 301 tggtcctaaa acccacttgc actcatacgc agggccctct gcagctcgag tctagcgacg 361 cgtgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc 421 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc 481 gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 541 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 601 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 661 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 721 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 781 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 841 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 901 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 961 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 1021 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 1081 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 1141 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 1201 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 1261 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 1321 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 1381 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 1441 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 1501 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 1561 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 1621 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 1681 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 1741 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 1801 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 1861 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 1921 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 1981 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 2041 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 2101 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 2161 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 2221 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 2281 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 2341 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg 2401 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg 2461 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct 2521 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa attgtaaacg 2581 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 2641 agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata gagttgagtg 2701 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 2761 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt 2821 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 2881 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 2941 gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 3001 ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt gtgaaatacc 3061 gcacagatgc gtaaggagaa aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg 3121 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc 3181 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg 3241 cggccgctcg gtccgcacgt ggttacctac aaaatcagaa ggacagggaa gggagcagtg 3301 gttcacgcct gtaatcccag caatttggga ggccaaggtg ggtagatcac ctgagattag 3361 gagttggaga ccagcctggc caatatggtg aaaccccgtc tctaccaaaa aaacaaaaat 3421 tagctgagcc tggtcatgca tgcctggaat cccaacaact cgggaggctg aggcaggaga 3481 atcgcttgaa cccaggaggc ggagattgca gtgagccaag attgtgccac tgcactccag 3541 cttggttccc aatagacccc gcaggcccta caggttgtct tcccaacttg ccccttgctc 3601 cataccaccc ccctccaccc cataatatta tagaaggaca cctagtcaga caaaatgatg 3661 caacttaatt ttattaggac aaggctggtg ggcactggag tggcaacttc cagggccagg 3721 agaggcactg gggaggggtc acagggatgc cacccgtaga tcatctctag actcgaggtc 3781 gaggccagag aggccatcga atggcgcgcc gcgatcgcgt ctcgaggtcg acggtatcga 3841 tgcggggagg cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga 3901 agaggccgca gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa 3961 gggacgtagc agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa 4021 ggaaaggacg atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc 4081 cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg 4141 aaagcgaaag tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt 4201 gcgtcagcaa acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca 4261 taaagagaca gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca 4321 atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag 4381 ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atcgataagc 4441 ttttgctcga agtcgaccta caggaacagg tggtggcggc cctcggagcg ctcgtactgt 4501 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca gcttggtgtc cacgtagtag 4561 tagccgggca gctgcacggg cttcttggcc atgtagatgg tcttgaactc caccaggtag 4621 tggccgccgt ccttcagctt cagggcctgg tggatctcgc ccttcagcac gccgtcgcgg 4681 gggtacaggc gctcggtgga ggcctcccag cccatggtct tcttctgcat tacggggccg 4741 tcggggggga agttggtgcc gcggaacttc accttgtaga tcagcgtgcc gtcctgcagg 4801 gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac 4861 ttgaagccct cggggaagga cagcttcttg tagtcgggga tgtcggcggg gtgcttcacg 4921 tacgccttgg agccgtactg gaactggggg gacaggatgt cccaggcgaa gggcaggggg 4981 ccgcccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc 5041 tcgccctcga tctcgaactc gtggccgttc acggagccct ccatgcgcac cttgaagcgc 5101 atgaactctt tgatgacgtc ctcggaggag gcggtgccgg agctgccgct gccggtgctg 5161 ccggtgccat gccccaggaa caggtggtgg cggccctcgg agcgctcgta ctgttccacg 5221 atggtgtagt cctcgttgtg ggaggtgatg tccagcttgg tgtccacgta gtagtagccg 5281 ggcagctgca cgggcttctt ggccatgtag atggtcttga actccaccag gtagtggccg 5341 ccgtccttca gcttcagggc ctggtggatc tcgcccttca gcacgccgtc gcgggggtac 5401 aggcgctcgg tggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcgggg 5461 gggaagttgg tgccgcggaa cttcaccttg tagatcagcg tgccgtcctg cagggaggag 5521 tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag 5581 ccctcgggga aggacagctt cttgtagtcg gggatgtcgg cggggtgctt cacgtacgcc 5641 ttggagccgt actggaactg gggggacagg atgtcccagg cgaagggcag ggggccgccc 5701 ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc 5761 tcgatctcga actcgtggcc gttcacggag ccctccatgc gcaccttgaa gcgcatgaac 5821 tctttgatga cgtcctcgga ggaggccacc atggcggccg cggatcccgg tgcagggccg 5881 ggattctcct ccacgtcacc gcatgttaga agacttcctc tgccctcggc cgcgggtacc 5941 gactttctag cggtcaccca atatgtctgc atttttccct ttcctttcac ttccatcagg 6001 tccagtggct gggtcacgaa ttttcccttg atcatctggt aagtagagtc ggagatatga 6061 atgtgccctg ccttgctggt tgactccatc cgggaagctg tattgacggt atcccccacc 6121 aggcaccaat gagggttcag gtcgcccagg actccggctg tcactggtcc actattcagg 6181 ccgatgcgaa tgttgataga ttcgccggta gcagttttaa aagtcttgat catctcgatg 6241 atgtccaggg cgaagttcac tgctcgatca gcgtggtcgg ggaccacctc tggtgctccg 6301 gtcactccca gataagcgtc cccaatagtt tcgaccttat acacgcccca ccgcttagcg 6361 atggcatcat actccagcca cagcttgttc agggtggcca tcatgtcctt tgtggaggtc 6421 ctggagctaa tcacagtaaa gtttgtgatg tcagaaaaga agactgtcac ggattcgtac 6481 tctttggcct cggttgcctg tccattcttc agctgttcca gcaccccctc aggcatcatc 6541 ttctgcagaa tggcgtcaca gttagtgatc tgagattcca gttcctctgc aatagctgtg 6601 atggcttcga ctttctgatt ctgtgcctgc tccacagtgc tgttgaccag caccagtgtc 6661 aggaacacct tggccccaat atcgatcatg gtaaaagatg cctcagtcat tgtgaaactc 6721 agcatgccag atgtaaaggc gaaccagctc agtggaaaca gggtccatgt ggtgacagtt 6781 gagaagcgtg tccacctcag gccattcact tcaatgttgc aaggggtctc tccgttgatg 6841 gccccagtaa aggacctcca caggctgtag accacgaagc taaagaaggc acatgacagg 6901 atattaaaca ggtttcccca gggataaggg ggcagcacag ctccaaagaa cccgcagacc 6961 accagggcgt aatcagagaa gaccaccccc caggcattat ggtctgcctg tgtaatctcg 7021 gagatcagca gaatcaggac gggacaggta gcaatccact ccaggtatct tgcaggctcc 7081 actgggttgc tcagggtgcc ttcgatggtt ggagtcagcc gcagcataat caggatgtat 7141 gaactaaaag cgatgatgtt cactcccagc aggcacaggg acagtgcttt cttctctggg 7201 tttctggcgc ctgcaaatcc catataactg aaagcaatcc cgatgccaaa tccgaaggcc 7261 ccatagccgt accaaatgtc tgcccagccc ctattataag cctgtctttc ctggataaac 7321 agagcgtaga aagcgccggc cacggtcaga atggcccaca tgataaactg tcctgtcagg 7381 aatttcagtg aaggggtact cagcatcata tcccaggacc acatgtctgc gatgtttgtc 7441 tggtactgct gcagagcgga ctgcttacta ggagagccag gtcggctatt ggcgggagag 7501 gacacgttgc ttgaacttgc agaggaggtg ctgcggacat tggacacgct cagtgaattg 7561 ttctgtctgt tccgcatatt agcgttgctt gaggcggtac tagagccagt gaccttgata 7621 ttgggctgac tgggagatgg cagcagtgag gctccggagc tgcggaaact tccgtttcga 7681 gtcagtggct gctcgacgat agggtgtttt gtcacgctac acttgcaatc gtcgcacagg 7741 gttgaggttg gtgagtaaca gtattcaggg caggaacagc cactacatgc tccccgcagg 7801 ttattatctt tatctttcat agacatagac attattgcga attcgactgc gctctcaggc 7861 acgacacgac tcctccgctg cccaccgcag actgaggcag cgctgagtcg ccggcgccgc 7921 agcgcagatg gtcgcgcccg tgccccccta tctcgcgcct cgcgtggtgc g //