LOCUS Exported 6338 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION express the genetically-encoded fluorescent dopamine(DA) sensor GRAB_DA1h in neurons. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-GRAB_DA1h SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6338) AUTHORS Sun F, Zeng J, Jing M, Zhou J, Feng J, Owen SF, Luo Y, Li F, Wang H, Yamaguchi T, Yong Z, Gao Y, Peng W, Wang L, Zhang S, Du J, Lin D, Xu M, Kreitzer AC, Cui G, Li Y TITLE A Genetically Encoded Fluorescent Sensor Enables Rapid and Specific Detection of Dopamine in Flies, Fish, and Mice. JOURNAL Cell. 2018 Jul 12;174(2):481-496.e19. doi: 10.1016/j.cell.2018.06.042. PUBMED 30007419 REFERENCE 2 (bases 1 to 6338) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6338 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 581..590 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide 587..649 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader protein_bind 650..674 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 1648..1669 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1775..1796) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2036..2055) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind complement(2518..2538) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2936..2947) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3085..3561 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3284..3303) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3601..3741 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3601..3730 /label=AAV2 ITR rep_origin 3816..4271 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3903..3922) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4113..4134 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4288..4307) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4407..4429 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4467..4485) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4553..4657 /gene="bla" /label=AmpR promoter CDS 4658..5518 /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(4876..4895) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5689..6277 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6178..6197 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac 181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag 241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg 301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg 361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc 421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc 481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga 541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac 601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta 661 caaaaaagtt ggcaccaccg gtgatccact gaatctgtcc tggtatgatg atgatctgga 721 gaggcagaac tggagccggc ccttcaacgg gtcagacggg aaggcggaca gaccccacta 781 caactactat gccacactgc tcaccctgct catcgctgtc atcgtcttcg gcaacgtgct 841 ggtgtgcatg gctgtgtccc gcgagaaggc gctgcagacc accaccaact acctgatcgt 901 cagcctcgca gtggccgacc tcctcgtcgc cacactggtc atgccctggg ttgtctacct 961 ggaggtggta ggtgagtgga aattcagcag gattcactgt gacatcttcg tcactctgga 1021 cgtcatgatg tgcacggcga gcatcctgaa cttgtgtgcc atcagcatcg acaggtacac 1081 agctgtggcc atgcccatgc tgtacaatac gcgctacagc tccaagcgcc gggtcaccgt 1141 catgatctcc atcgtctggg tcctgtcctt caccatctcc tgcccactcc tcttcggact 1201 caataacgca gaccagaacg agtgcatcat tgccaacccg gccttcgtgg tctactcctc 1261 catcgtctcc ttctacgtgc ccttcattgt catgctgctg gtctacatca agatctacat 1321 tgtcctccgc agacgccgca agcgagtcaa caccaaacgc agcagccgag ctttcagggc 1381 ccacctgagg gctccactaa agggcaactg tactcacccc gaggacatga aactcggagg 1441 aaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact tccacatccg 1501 ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat 1561 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt ccaaactttc 1621 gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 1681 gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca tggtgagcaa 1741 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa 1801 cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac 1861 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac 1921 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt 1981 cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga 2041 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat 2101 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta 2161 caacactgga gcagcagcaa tgagccgtag gaagctctcc cagcagaagg agaagaaagc 2221 cactcagatg ctcgccattg ttctcggcgt gttcatcatc tgctggctgc ccttcttcat 2281 cacacacatc ctgaacatac actgtgactg caacatcccg cctgtcctgt acagcgcctt 2341 cacgtggctg ggctatgtca acagcgccgt gaaccccatc atctacacca ccttcaacat 2401 tgagttccgc aaggccttcc tgaagatcct ccactgctga gaattcgata tcaagcttat 2461 cgatcatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 2521 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 2581 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 2641 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 2701 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 2761 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 2821 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 2881 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 2941 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 3001 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata 3061 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc 3121 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc 3181 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg 3241 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc 3301 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt 3361 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa 3421 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc 3481 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc 3541 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc 3601 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 3661 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 3721 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg 3781 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag 3841 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc 3901 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 3961 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4021 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4081 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4141 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta 4201 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac 4261 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 4321 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc 4381 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc 4441 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt 4501 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac 4561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 4621 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 4681 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 4741 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 4801 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 4861 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 4921 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 4981 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 5041 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 5101 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 5161 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 5221 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 5281 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 5341 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 5401 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 5461 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 5521 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 5581 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 5641 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 5701 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 5761 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 5821 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 5881 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 5941 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 6001 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 6061 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 6121 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 6181 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 6241 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 6301 acggttcctg gccttttgct ggccttttgc tcacatgt //