LOCUS Exported 6482 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses the genetically-encoded fluorescent norepinephrine(NE) sensor GRAB_NE1m in neurons. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-GRAB_NE1m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6482) AUTHORS Feng J, Zhang C, Lischinsky JE, Jing M, Zhou J, Wang H, Zhang Y, Dong A, Wu Z, Wu H, Chen W, Zhang P, Zou J, Hires SA, Zhu JJ, Cui G, Lin D, Du J, Li Y TITLE A Genetically Encoded Fluorescent Sensor for Rapid and Specific In Vivo Detection of Norepinephrine. JOURNAL Neuron. 2019 May 22;102(4):745-761.e8. doi: 10.1016/j.neuron.2019.02.037. Epub 2019 Mar 25. PUBMED 30922875 REFERENCE 2 (bases 1 to 6482) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6482 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 74..203 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 654..663 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide 660..722 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader protein_bind 723..747 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind 1850..1871 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1977..1998) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2238..2257) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind complement(2735..2755) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3153..3164) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3302..3778 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3501..3520) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3818..3958 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3818..3947 /label=AAV2 ITR rep_origin 4033..4488 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4120..4139) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4330..4351 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4505..4524) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4624..4646 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4684..4702) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4770..4874 /gene="bla" /label=AmpR promoter CDS 4875..5735 /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(5093..5112) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5906..12 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6395..6414 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 61 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg 121 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc 181 caactccatc actaggggtt cctgcggccg cacgcgtatg aggcggggtg ggggtgccta 241 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca 301 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc 361 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag 421 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct 481 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg 541 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc 601 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggtgccacca 661 tggagacaga cacactcctg ctatgggtac tgctgctctg ggttccaggt tccactggtg 721 acacaagttt gtacaaaaaa gttggcacca ccggtggctc cctgcagccg gacgcgggca 781 acgcgagctg gaacgggacc gaggcgccgg ggggcggcgc ccgggccacc ccttactccc 841 tgcaggtgac gctgacgctg gtgtgcctgg ccggcctgct catgctgctc accgtgttcg 901 gcaacgtgct cgtcatcatc gccgtgttca cgagccgcgc gctcaaggcg ccccaaaacc 961 tcttcctggt gtctctggcc tcggccgaca tcctggtggc cacgctcgtc atccctttct 1021 cgctggccaa cgaggtcatg ggctactggt acttcggcaa ggcttggtgc gagatctacc 1081 tggcgctcga cgtgctcttc tgcacgtcgt ccatcgtgca cctgtgcgcc atcagcctgg 1141 accgctactg gtccatcaca caggccatcg agtacaacct gaagcgcacg ccgcgccgca 1201 tcaaggccat catcatcacc gtgtgggtca tctcggccgt catctccttc ccgccgctca 1261 tctccatcga gaagaagggc ggcggcggcg gcccgcagcc ggccgagccg cgctgcgaga 1321 tcaacgacca gaagtggtac gtcatctcgt cgtgcatcgg ctccttcttc gctccctgcc 1381 tcatcatgat cctggtctac gtgcgcatct accagatcgc caagcgtcgc acccgcgtgc 1441 cacccagccg ccggggtccg gacgccgtcg ccgcgccgcc ggggggcacc gagcgcaggc 1501 ccaacggtct gggccccgag cgcagcgcgg gcccgggggg cgcagaggcc gaaccgctgc 1561 ccacccagct caacggcgcc cctggcgagc ccgcgccggc cgggccgcgc gacaccgacg 1621 cgctggacct ggaggaggga ggaaacgtct atatcaaggc cgacaagcag aagaacggca 1681 tcaaggcgaa cttccacatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc 1741 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 1801 tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc 1861 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta 1921 ccggagggag catggtgaga aagggcgagg agctgttcac cggggtggtg cccatcctgg 1981 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggtgagggcg 2041 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc 2101 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg 2161 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc 2221 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 2281 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 2341 tcctggggca caagctggag tacaacaccg gagcagcagc acgctggcgc gggcggcaga 2401 accgcgagaa gcgcttcacg ttcgtgctgg ccgtggtcat cggagtgttc gtggtgtgct 2461 ggttcccctt cttcttcacc tacacgctca cggccgtcgg gtgctccgtg ccacgcacgc 2521 tcttcaaatt cttcttctgg ttcggctact gcaacagctc gttgaacccg gtcatctaca 2581 ccatcttcaa ccacgatttc cgccgcgcct tcaagaagat cctctgtcgg ggggacagga 2641 agcggatcgt gttgtgagaa ttcgatatca agcttatcga tcatcaacct ctggattaca 2701 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 2761 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 2821 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 2881 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 2941 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 3001 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 3061 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga 3121 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 3181 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 3241 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta 3301 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc 3361 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc 3421 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac 3481 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg 3541 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt 3601 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg 3661 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc 3721 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct 3781 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg 3841 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga 3901 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg 3961 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa 4021 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 4081 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt 4141 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 4201 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt 4261 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 4321 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt 4381 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt 4441 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc 4501 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg 4561 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg 4621 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa 4681 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga 4741 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 4801 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 4861 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 4921 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 4981 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 5041 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 5101 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 5161 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 5221 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 5281 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 5341 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 5401 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 5461 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 5521 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 5581 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 5641 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 5701 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 5761 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 5821 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 5881 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 5941 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 6001 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 6061 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 6121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 6181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 6241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 6301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 6361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6481 gg //